SWIFS FBU Serology Procedures Manual v2.0 (02.04.2009) - Re Formatted

Southwestern Institute of Forensic Sciences Criminal Investigation Laboratory
Forensic Biology Unit Serology Procedures Manual, Version 2.0 (2.4.2009)
Approved by: Stacy McDonald, Ph.D., Deputy Section Chief Timothy J. Sliter, Ph.D., Section Chief Karen Young, Quality Manager
Serology Procedures Manual (2.4.2009)
1. Title Page 2. Table of Contents 3. Corrections & Revisions Log 4. Background 5. Brentamine Fast Blue B Test for Acid Phosphatase 6. Microscopic Examination of Smears for Spermatozoa 7. Screening Evidence with an Alternate Light Source 8. Collection and Storage of Test Areas 9. OneStep ABAcard p30 Test for the Identification of Semen 10. Leucomalachite Green (LMG) Test for Blood 11. OneStep ABAcard HemaTrace for the Forensic Identification of Human Blood 12. Species of Origin Determination Using the Ouchterlony Double Diffusion Method 13. Buccal Swab Storage 14. Sexual Assault Clothing Evidence Analysis 15. Blood Evidence Analysis 16. Evidence Handling and Analysis Procedures for Sexual Activity Kits 17. Examination and Processing of Hair Combings 18. Collection, Preservation, and Documentation of Trace Evidence 19. Storage of Evidence in the Forensic Biology Freezer 20. Post-conviction Case Research Guidelines 21. Appendix 1: Reagents and Solutions: Preparation, Storage, and QC 22. Appendix 2: Abbreviations for Bench Notes 23. Appendix 3: Critical Reagents List 24. Appendix 4: Quality Control for Serological Examination and Sample Processing 25. Appendix 5: Worksheets
Serology Procedures Manual (2.4.2009) Table of Contents
Corrections & Revisions (2.4.2009) Serology Procedures Manual Effective Date 8/16/2002 12/11/2002 Authorizing Individual Sliter Sliter
1/16/2003 7/3/2003 1/24/2008
2/4/2009
Description Deletion: Acid Phosphatase Quantitative Assay Deletion: P30 Crossover Electrophoresis Test Addition: Onestep ABAcard P30 Test for the Identification of Semen Addition: Post-conviction Case Research Guidelines Addition: Onestep ABAcard HemaTrace Test for the Forensic Identification of Human Blood Individual pdf procedure files were consolidated into a single pdf collection document. No content change to the individual procedures. Changes from Version 1.0 to Version 2.0 Conversion of various procedures from images to electronic text documents. Incorporation of procedures documented in memoranda into the manual. Revision of procedures to improve clarity. Various insubstantial changes made (e.g., removal of references to individual page numbers). Significant changes: Table of Contents removal of page numbers and incorporation of procedures previously documented in memoranda Screening Evidence Using An Alternate Light Source Removal of procedure for the Omnichrome 1000. OneStep ABAcard p30 Test for the Identification of Semen Revision of procedure to improve clarity. New procedure for the documentation of results. OneStep ABAcard HemaTrace for the Forensic Identification of Human Blood Revision of procedure to improve clarity. New procedure for the documentation of results. Sexual Assault Clothing Evidence Analysis Revision of procedure to improve clarity and to provide references to p30 and HemaTrace tests. Blood Evidence Analysis References to HemaTrace test added to procedure.
Sliter Sliter Sliter
McDonald
Serology Procedures Manual Corrections & Revisions Log (2.4.2009)
Evidence Handling and Analysis Procedures for Sexual Activity Kits Revision of procedure to reflect current practice of the laboratory and to improve clarity. Appendix 1: Reagents and Solutions: Preparation, Storage, and QC Removal of QC procedures and reagents not currently used by the laboratory. Entire QC procedure for HemaTrace revised to reflect the current practices of the laboratory and to provide additional guidance. Appendix 3: Abbreviations for Bench Notes Additional abbreviations added. Appendix 5: Worksheets Additional worksheets added. Worksheets not currently used by the laboratory were removed.
Serology Procedures Manual Corrections & Revisions Log (2.4.2009)
BACKGROUND ON SEROLOGICAL TESTING OF FORENSIC EVIDENCE (2.4.2009) PRINCIPLE Serological analysis of evidence is performed in order to identify bodily fluids that may be used for human identification. Analysis of evidence may include examination of physical evidence collected from the complainant during a medical examination or autopsy, any clothing worn by either suspect or victim during the alleged assault, or other evidence collected from the crime scene. Initially, clothing or other items may be examined with an alternate light source in order to identify areas which may contain biological fluids such as semen. Analysis is performed through presumptive chemical tests for the presence of seminal fluid and/or for the presence of blood. Confirmatory tests for semen include microscopic examination for spermatozoa, and the P30 test which identifies a seminal fluid specific protein. The presence of human blood is confirmed using an antibody specific for human blood. Documentation of findings, collection of samples from the submitted evidence, and frozen storage of test samples are performed in the event that further analysis is needed. I. Purpose of Serology Procedures Manual The following procedures are designed to cover routine casework circumstances encountered in the Forensic Biology (FB) Laboratory. Modification of these procedures and/or policies may be required under specific case circumstances. Any alteration of these procedures and/or policies must adhere to the guidelines established in the FB Quality Management Program. II. General Quality Control All procedures in this manual must conform to Physical Evidence Section and FB Quality Control Guidelines described in detail in other documents. Refer to Appendix 4, for more information. III. General Safety All procedures must be performed in compliance with Institute safety policies. Consult the Institute Biological Exposure Plan and the Hazardous Chemical and Safety Manual for more information.
Serology Procedures Manual BACKGROUND ON SEROLOGICAL TESTING OF FORENSIC EVIDENCE (2.4.2009)
BRENTAMINE FAST BLUE B TEST FOR ACID PHOSPHATASE (2.4.2009) PRINCIPLE Acid Phosphatase (ACP) is an enzyme present at high levels in seminal fluid. Swab analysis involves the application of a qualitative colorimetric test called the Brentamine Fast Blue B test. Using specific substrates, this test provides a rapid visual screening of swabs that may contain the ACP enzyme. It has been determined that ACP removes ester-linked phosphate groups from monophosphate substrates through hydrolysis. Therefore, it is practical to use a substrate that can easily undergo this reaction. This method uses alpha-naphthyl phosphoric acid (an artificial substrate) reacted with a diazonium compound to form an insoluble colored product. However, this test does not specifically identify seminal fluid, since the ACP enzyme is also present in other bodily fluids as well as other organisms; it is only a presumptive test for seminal fluid. I. Swab analysis using the transfer method Analysis of vaginal, anal, and oral swabs collected from the complainant and submitted in the sexual activity kit is performed to detect the presence of the enzyme, acid phosphatase (ACP). The transfer method is used to perform the Brentamine Fast Blue B presumptive test while preserving the swab containing the original sample. A. Using forceps cleaned with a 10% bleach solution, place a Whatman paper square in the well of a clean spot plate. B. Using a clean pipette, place one drop of Millipore water on the Whatman paper to moisten it. C. Firmly roll the sides and the tip of the swab to be tested on the paper. This transfers a small amount of material from the swab to the paper. D. Pipette one to two drops of Brentamine reagent onto the moistened paper which contains the transferred sample. E. Allow 15 seconds to pass while observing for the presence or absence of a color change to purple/pink on the paper. Record the test results. F. Discard the paper squares. Allow swabs to dry. G. Place positive swabs into a baggie labeled with the unique Forensic Laboratory Number (FL#), and item #. i. Place the baggie in a plastic envelope labeled with the FL#, item #(s), complainants name, analysts initials, and storage date. ii. Heat seal the envelope. iii. Date and initial the seal. iv. File the envelopes in the evidence storage freezer. v. Follow proper procedure concerning documentation of evidence transfer on the internal chain (see Storage of Evidence in the Forensic Biology Freezer).
Serology Procedures Manual BRENTAMINE FAST BLUE B TEST FOR ACID PHOSPHATASE (2.4.2009)
II. Swab analysis using the direct testing method Clothing, other large items, or other evidence may be examined for the presence of ACP by sampling specific stains or areas on the item with a sterile cotton swab and directly performing the Brentamine Fast Blue B test on the swab. A. Obtain several sterile cotton swabs and using a minimal amount of deionized water, lightly moisten the cotton ends of the swabs. B. Sample the questioned area (e.g. visible stain and/or ALS positive area) by firmly swabbing with a moistened swab. C. Perform the ACP test by applying one to two drops of the Brentamine reagent on the swab and wait 15 seconds for a result (See Collection and Storage of Test Areas). D. If apparent blood stains are observed on the questioned area, immediately perform the LMG test on the same swab using the procedure, LMG Test for Blood. Note: Both swab analysis methods may be used to test for blood instead of seminal fluid by performing the LMG test for blood instead of the Brentamine test. III. Interpretation of Results A. The presence of a color change from white (paper or swab color) to purple/pink indicates that a reaction has taken place. i. If no color change to purple/pink has occurred within 15 seconds, the results are recorded as negative. ii. If a color change to purple/pink is very faint within the 15 seconds then the results are to be considered a weak reaction and recorded as weak positive. iii. If there is a color change to purple/pink then the results are to be considered positive and are recorded as positive. B. Record the results on the SA kit or Evidence Examination worksheet. For example:
ACP x 4 * ACP x 4 * ACP wk x 2*

* = x4 or x2 refers to the number of swabs tested that gave the indicated results. Recording the number is not necessary for swabs used to sample questioned areas on evidence.
Serology Procedures Manual BRENTAMINE FAST BLUE B TEST FOR ACID PHOSPHATASE (2.4.2009)
MICROSCOPIC EXAMINATION OF SMEARS FOR SPERMATOZOA (2.4.2009) PRINCIPLE Microscopic analysis is performed to detect the presence of spermatozoa on smears obtained in the Sexual Activity (SA) kit, or prepared from evidentiary samples. Southwestern Institute of Forensic Sciences (S.W.I.F.S.) Forensic Biology Laboratory employs a staining procedure involving Nuclear Fast Red and Picroindigocarmine stain. This is also called the Christmas tree stain. Using this stain, spermatozoa heads are stained red and the spermatozoa tails are stained green. I. Christmas Tree Staining Procedure A. Using a pencil, label the slide with the FL#, the item number, last name, and first initial of the complainant. B. Spray the cell side of the smear with cytological fixative and allow to dry. C. Place smear in a staining tray cell side up. D. Cover the slide with Nuclear Fast Red solution and let stand for at least 20 minutes. E. Rinse the slide with distilled water. F. Cover the slide with Picroindigocarmine and let stand for 15-30 seconds. G. Rinse the slide using 95% ethanol until the excess stain is cleaned off. H. Allow the smear to dry before microscopic examination. I. Smears may be re-stained as necessary.
II. Interpretation of Results A. Results are recorded on the SA Kit or Evidence Examination worksheet after microscopic analysis is completed. i. If no spermatozoa are observed, record the results as negative (). ii. Date and initial the results. iii. Negative and rare slides must be verified by another analyst. iv. Coordinates of rare spermatozoa should be noted. B. If spermatozoa are seen on the smear, the spermatozoa are to be described as intact (head and tail) or heads only. i. The following guidelines are used to approximate the number of spermatozoa seen on the slide: Term used Rare Occasional Few Moderate Many Approximate number of sperm cells 1-7 per slide 15-20 per slide Greater than 20 per slide Two per field on average 3-4 per field on average
Serology Procedures Manual MICROSCOPIC EXAMINATION OF SMEARS FOR SPERMATOZOA (2.4.2009)
ii.
The following are examples of how to record the results: Quantity Description Rare heads Many heads/many intact
Coordinates 35.9 x 100.1
Initials and Date KRD 5/9/08 KRD 5/9/08
Serology Procedures Manual MICROSCOPIC EXAMINATION OF SMEARS FOR SPERMATOZOA (2.4.2009)
SCREENING EVIDENCE USING AN ALTERNATE LIGHT SOURCE (2.4.2009) PRINCIPLE S.W.I.F.S. currently employs the CrimeScope CS-55 Forensic Light Source to screen evidence. It is useful to examine sexual assault clothing or other evidence using a wavelength of 445-450 nm because biological fluids such as seminal fluid, saliva, and urine are known to fluoresce at this wavelength. I. Procedure for use of the CrimeScope CS-55 Forensic Light Source A. B. C. D. E. Place the FORENSIC LIGHT SOURCE IN USE sign outside the door. Turn on the fan from the switch at the back of the CrimeScope, then turn on the light. Turn Wheel 2 to Max Power setting or other appropriate open shutter setting. Clean bench table with fresh 10% bleach solution and lay down a clean sheet of white table paper. Open the packaged item with a scalpel, if necessary. Make sure not to break any other seals on the packaging in which the item was received, unless absolutely necessary (follow Institute Evidence Handling procedures described in detail in other documents). Place the item neatly on the clean sheet of white table paper. Wear ultraviolet light protective goggles (yellow in color) before adjusting wavelength and while examining evidence items. All lights in the immediate area should be turned off when using the CrimeScope to detect stains. Examine the item at 445 nm by adjusting the wavelength using either the buttons on the handheld lightsource, or Wheel 1 (knob on front of the CrimeScope). At this point the wavelength can be fine tuned using the +/- knob on the front of the instrument. Slowly sweep the light source wand over the entire item without touching the item with the wand. i. Examine the front, back, inside, and outside of the item including the inside crotch of shorts and pants. An ultraviolet (UV) light positive stain will fluoresce a greenish color. Note any UV light positive stain by encircling the area using a permanent ink marker. Turn off the instrument by turning off the light and then the fan after cooling. Record usage in the CrimeScope Log.
F. G. H. I.
J.
K. L. M. N.
II. Documentation Questioned areas identified using an alternate light source may be subjected to further analysis (e.g. Brentamine test), as necessary. These areas should be identified as Omni positive (Omni ) on the Evidence Examination or SA kit worksheet. Further information regarding Omni positive areas is given in the Sexual Assault Clothing Evidence Analysis Procedure.
Serology Procedures Manual SCREENING EVIDENCE USING AN ALTERNATE LIGHT SOURCE (2.4.2009)
COLLECTION AND STORAGE OF TEST AREAS (2.4.2009) PRINCIPLE When a specific area on an item tests positive using either a presumptive or a confirmatory test, representative samples are to be removed and preserved in the event that further analysis may be performed on the evidence. Additionally, samples may be collected from an item for the purpose of DNA analysis only and without serological testing (e.g. to determine wearer/handler). I. Procedure for taking a cutting from an item A. Use 10% bleach solution to decontaminate scissors and forceps. Be sure to wipe utensils with gauze to make sure that the utensils are not wet. B. Cut out the positive area of the item using the clean scissors and forceps or a sterile scalpel. 1. The entire stain or area need not be retained; however, a portion should be retained that is large enough for both further analysis by S.W.I.F.S., and for storage of an additional portion for analysis, if requested. 2. Place the test area (positive area) into a baggie labeled with the unique Forensic Laboratory Number (FL#), item #, and test area number (e.g. T 1 ). 3. Place the baggie in a plastic envelope labeled with the FL#, item #s, complainants name, analysts initials, and storage date. 4. Heat seal the envelope. 5. Date and initial the seal. 6. File the envelopes in the evidence storage freezer. 7. Follow proper procedure concerning documentation of evidence transfer on the internal chain (see Storage of Evidence in the Forensic Biology Freezer). II. Procedure for collecting a swabbing of item A. Some samples may be retained by swabbing with a sterile cotton swab rather than removing a cutting. 1. This method is preferable on items which are difficult to cut, or when the stain appears to be topical rather than soaked in. 2. This technique may also be used on larger areas, or areas which are to be analyzed for DNA only (e.g. for wearer/handler information). 3. Under certain case circumstances, an attempt will be made to collect more than one swab in case additional testing is requested at a later time. 4. Allow the swab(s) to dry before storing. 5. Follow same storage procedures as described for cuttings (see above). B. Using a permanent marker, label the area that was retained or swabbed as appropriate (e.g. T 1 ), on the item for identification purposes. If it is not possible to label the test area on the item, this information must be noted on the worksheet. III. Procedure for the retention of entire item
Serology Procedures Manual COLLECTION AND STORAGE OF TEST AREAS (2.4.2009)
A. Occasionally it will be necessary to retain the entire item submitted for analysis (e.g. swab, swatch of cloth, or dirt sample). 1. Store entire item following the same storage procedures previously listed. 1. Write FL#, item#, initials, and date on the original packaging. 2. Seal the packaging with evidence tape and initial the seal. 3. Note on the SA kit analysis or Evidence examination worksheet that the entire item has been retained. 4. Indicate on the internal chain that packaging only is being returned to the proper agency. IV. Documentation Document all test areas or items retained on the Evidence Examination or SA kit worksheet. See Sexual Assault Clothing Evidence Analysis for more information.
Serology Procedures Manual COLLECTION AND STORAGE OF TEST AREAS (2.4.2009)
OneStep ABAcard p30 Test for the Identification of Semen (2.4.2009) PRINCIPLE This document describes the procedures for the confirmatory test for semen based upon detection of the prostate specific antigen p30 using the OneStep ABAcard p30 Test (Abacus Diagnostics). Dried stains are solubilized in water, and the extract is applied to the sample well of the p30 test device. If p30 is present, it binds with a dye-conjugated anti-human p30 antibody and forms a mobile antigen-antibody-dye complex. This complex then migrates through the device towards the test area designated T. In the test area, a second anti-human p30 antibody is immobilized. The immobilized antibody captures the mobile complex, resulting in concentration of the conjugated dye in a narrow zone on the membrane. If the p30 concentration in the extract exceeds 4 ng/mL, a pink line forms in the test area, indicating a positive result. As an internal process control, free antibody-dye conjugate (i.e., conjugate not bound to p30) are captured by an immobilized anti-immunoglobulin antibody located in the control area designed C, resulting in a pink line. Because the antibody-dye conjugate is normally at a molar excess compared to the p30 in the extract, the control line is expected to appear and indicates proper performance of the test device in the absence of a line being present in the test area. REAGENTS, SUPPLIES & MATERIALS 1. 2. 3. 4. Spin baskets and matching tubes (LPS Cat. # MB50092 or equivalent) Seminal Fluid Standard 1:25,000 (prepared from SERI R563 Semen Standard) ABAcard p30 Test Kit (Abacus Diagnostics p/n 308332) Deionized water
SAMPLE REQUIREMENTS Samples will consist of portions of fabric, swabs, and other materials that may contain semen/seminal fluid based. Typically, the need for p30 testing by this procedure will be indicated by previous presumptive chemical testing. Indications other than presumptive chemical testing may apply in specific case circumstances. WORKSHEETS 1. ABAcard P30 Worksheet PROCEDURE NOTE: Before beginning this procedure, ensure that all critical reagents, solutions and materials have been QCed and have not passed their expiration date. NOTE: Test results obtained with the p30 test devices must be observed and confirmed by a qualified analyst. A. Sample Extraction
Serology Procedures Manual OneStep ABAcard p30 Test for the Identification of Semen (2.4.2009)
1. Place the test sample for extraction in the extraction tube. This tube must fit the spin basket that is subsequently used. a. For intimate swabs, the following general guidelines are to be used in determining the amount of sample to be tested. i. For a swab that gives a strong presumptive ACP reaction, or less of a swab may be used. ii. For a swab that gives a weak presumptive ACP reaction, up to of a swab may be used. iii. The maximum amount that can be used for p30 testing is of a swab, so as to preserve sufficient sample for DNA testing. b. For stains on fabric, use approximately 0.5 cm2. i. This amount may be adjusted based upon the intensity of the presumptive ACP reaction and the size of the stain. ii. It is important that sufficient sample be retained for DNA testing. 2. Add 250 L deionized water. a. Depending on the size, condition and texture of the substrate material being extracted, more than 250 L of water may be needed in some cases to fully submerse the material. b. The minimum amount of water needed to fully submerse the substrate should be used in these cases. 3. Fully saturate and submerge the substrate by centrifuging and/or vortexing. 4. Incubate a minimum of 2 hours at room temperature. a. The test sample may be incubated overnight at 4C. 5. Following incubation, vortex the tube to dislodge cellular material from the substrate. a. For large samples, vortexing the tube in an inverted position may be necessary to achieve sufficient agitation. 6. Centrifuge the tube for several seconds at 1,000 rpm to collect the liquid. 7. Transfer the substrate to a spin basket and place the spin basket in the tube. a. Transfer should be done using a clean implement, such as a sterile pipette tip or bleached forceps. b. Care must be taken to avoid cross-contamination from one tube to another within a batch. 8. Centrifuge the spin basket/tube for 1 minute on highest speed to recover liquid and cellular material from the substrate. 9. Discard the basket and the substrate. B. Spermatozoa Slide Preparation 1. Prepare a microscope slide for spermatozoa search by transferring the debris pellet in 10 L liquid to a labeled microscope slide. Allow the slide to air dry. 2. Follow the Christmas tree staining procedure as described in the Microscopic Examination of Smears for Spermatozoa protocol. 3. Save the remaining extract for the p30 test. The test may be performed immediately, or the extract may be stored, either at 4oC for up to 1 week, or indefinitely at -20oC. C. Testing for the p30 Antigen 1. The p30 test is performed in a batch-wise manner.
2. Each batch includes the following samples processed in parallel: a. Test Sample Extracts: Test sample extracts are prepared as described in Part A. 200 L of each extract is used in the p30 test. b. Positive Control: The positive control is 200 L of Seminal Fluid Standard 1:25,000. c. Negative Control: The negative control is 200 L of deionized water. The deionized water used as a negative control must be the same deionized water used to extract the test samples. 3. Allow any refrigerated samples to warm to room temperature before proceeding with the test. 4. Prepare an ABAcard p30 Worksheet for recording the results of the test. 5. Label the p30 test devices with the sample identifier. a. For test sample extracts, this will be the FL# & sample #. b. For controls, this will be POS (or ) or NEG (or ). 6. Initial and date each p30 test device. 7. Vortex the test sample extracts. 8. Transfer 200 L of the supernatant from each test sample into the sample well of the corresponding p30 test device. a. If several extracts are being processed in a batch-wise fashion, apply the extracts to the corresponding devices in quick succession, avoiding delays. 9. Transfer 200 L of the positive control (Seminal Fluid Standard, 1:25,000) to the corresponding p30 test device ( control). 10. Transfer 200 L of the negative control (deionized water) to the corresponding p30 test device ( control). 11. Incubate the test devices for 10 minutes at room temperature. 12. At the end of the 10 minute incubation, read the results of the tests by visual examination of the p30 test devices and record them on the results sheet (for further discussion of the interpretation and responses to test results, see Section D.). Abnormal or unusual test results that fall outside the following guidelines will be brought to the attention of a supervisor for evaluation on a case-by-case basis. a. Positive result. If there are 2 pink lines, one in the test area T and one in the control area C, then the test is Positive. b. Weak positive result. If there are 2 pink lines, one in the test area T and one in the control area C, and the T line is fainter than the T line on the positive control test, then the test is Weak Positive. c. Negative result. If there is a pink line in the control area C, but no line in the test area T, then the test is Negative. d. Inconclusive. If there is no pink line in the control area C, then the test is Inconclusive. 13. Immediately after reading the results, photograph the p30 test devices to document the test. a. Note: The digital photograph serves only to document the test. The test results are based upon direct visual examination of the test devices, not the photographs. Faint lines may not be visible in the hardcopy photographs. 14. Print the picture of the devices and attach it to the worksheet.
D. Evaluation of the p30 test results & responses to test results 1. Evaluate the negative control. a. The negative control is expected to give a negative result. b. If the negative control fails (i.e., gives a positive or inconclusive result), then the test is unreliable and must be repeated. 2. Evaluate the positive control. a. The positive control is expected to give a positive result. b. If the positive control fails (i.e., gives a negative or inconclusive result), then the test is unreliable and must be repeated. 3. Evaluate the results for the test samples. a. Positive result. i. Per the manufacturer, a positive result indicates that the p30 concentration in the extract is above 4ng/mL. ii. A positive result is reported as Positive. b. Weak positive result. i. A weak positive result may not be readily visible when the p30 test device is photographed. ii. A weak positive result is reported as Positive. c. Negative result. i. A negative result may indicate that: (a) p30 is not present in the extract (b) p30 is present at a level below the detection limit of the assay (c) p30 is present at an extremely high level, resulting in the High Dose Hook Effect. ii. To determine if the p30 is present at a high level, dilute the remaining extract 1:10 (20 L extract, 180 L water) and repeat test. Note: A microscope slide does not have to be prepared from the 1:10 dilution of the extract. (a) If the diluted sample gives a negative result, and if there is insufficient sample quantity to re-extract the sample, then the test result is Negative. (b) If the result is positive, then the test result is Positive. (c) If the diluted sample gives a negative result, and if there is sufficient sample quantity to re-extract the sample, then extract more of the sample and repeat assay. (i) If the result is positive, then the test result is Positive. (ii) If the result is again negative, then the test result is Negative. d. Inconclusive result i. If the test is inconclusive (due to absence of a pink band in the Control area), and if there is sufficient sample to repeat the test, then the test must be repeated. ii. If the test is inconclusive, and if there is insufficient sample to repeat the test, then the test result is Inconclusive. E. Documentation of test results. 1. The ABAcard p30 Worksheet is available as an electronic (.doc) form. a. Header and sample information may be filled in electronically. The form will then be printed.
b. Alternatively, the header and sample information may be filled in manually on a hardcopy printout of the form. 2. Test results are to be filled in manually on a hardcopy form. a. If the batch includes test samples from multiple cases, then one case packet will contain the original worksheet. The remaining case packets will contain a copy of the worksheet. 3. Photographs of p30 test devices are to be printed out and attached to the p30 worksheet. a. If there are samples from multiple cases in the batch, then print sufficient photographs for each report packet. b. Original photographs (not copies) must be included in each report packet. F. Reporting results and conclusions 1. Reports should clearly indicate the test results for both the p30 antigen test and the microscopic examination for spermatozoa. 2. E.g., The prostate specific protein p30 was [was not] detected on item 1. As part of the p30 test, a slide was prepared to screen for the presence of sperm cells. Sperm cells were [were not] detected on this slide.
LEUCHOMALACHITE GREEN (LMG) TEST FOR BLOOD (2.4.2009) PRINCIPLE Evidentiary items can be screened for blood by performing a presumptive chemical test using leuchomalachite green (LMG) reagent. This catalytic test is based on the peroxidase-like activity of the heme group. A color change is observed when LMG is oxidized in the presence of hemoglobin. This presumptive test for blood will routinely detect whole blood diluted 1:100 and has been successfully used in detecting blood at 1:10,000. I. LMG Reagent QC A. QC the LMG reagent and activator solutions before use. 1. Obtain a pair of LMG reagent and activator solutions, a positive control swab (human blood diluted 1:10,000), and a negative control swab (a sterile swab moistened with dI H 2 O). 2. Visually inspect the reagent and activator solutions. If either solution has a bluegreen appearance, discard the pair of solutions. Obtain an alternative pair of LMG reagent and activator solutions. 3. Add a drop of reagent solution, immediately followed by a drop of activator solution to each control swab. 4. A blue-green color change should be observed on the positive control swab within 5 seconds 5. If a color change is noted after addition of the LMG reagent solution, but before the addition of LMG activator, the reaction is a false-positive and the LMG solution should undergo the necessary QC to determine if it has expired. 6. No blue-green color change should be observed on the negative control swab. B. Record the LMG Lot #, date of QC, and initials on the SA Kit or Evidence Examination worksheet. 1. The LMG reagent pair must pass QC before proceeding with the analysis. 2. Each LMG reagent pair must pass QC daily before use. II. Analysis A. Sample the area using the swab analysis or other appropriate method. 1. Add a drop of reagent solution, immediately followed by a drop of activator solution to each swab. 2. Observe the swab for a color change. B. Record the results of analysis on the SA Kit or Evidence Examination worksheet. III. Interpretation of Results A. The immediate observation of a color change to blue-green on the swab after the addition of LMG activator solution indicates that a reaction has taken place. 1. If a color change occurs, the result is a positive presumptive test for blood and is recorded as CAT . (see Collection and Storage of Test Areas)
Serology Procedures Manual
LEUCHOMALACHITE GREEN (LMG) TEST FOR BLOOD (2.4.2009)
2. If the swab shows a color change in a small dot-like pattern, this is noted to be positive for traces of blood. These results are recorded as CAT traces. Traces may be noted as weak or strong if the analyst believes that additional clarification is necessary. 3. If a color change is noted after addition of the LMG reagent solution, but before the addition of LMG activator, the reaction is a false-positive and the LMG solution should undergo the necessary QC to determine if it has expired. B. In the event that there is no color change, the result is negative for blood and is recorded as CAT .

LEUCHOMALACHITE GREEN (LMG) TEST FOR BLOOD (2.4.2009)
OneStep ABAcard HemaTrace Test for the Forensic Identification of Human Blood (2.4.2009) PRINCIPLE This document describes the procedure for the confirmatory test for human blood based upon the detection of the human hemoglobin-specific antigen using the OneStep ABAcard HemaTrace Test (Abacus Diagnostics). Dried stains are solubilized in an extraction buffer, and the extract is applied to the sample well of the test device. If human hemoglobin is present, it will bind with a dye-conjugated anti-human hemoglobin antibody and form a mobile antigen-antibody-dye complex. This complex will then migrate through the device toward the test area designated T. In the test area, a second anti-human hemoglobin antibody is immobilized. The immobilized antibody captures the mobile complex resulting in concentration of the conjugated dye in a narrow zone on the membrane. When the human hemoglobin concentration exceeds 0.05 g/ml, a pink line forms in the test area indicating a positive result. As an internal process control, mobile antibody-dye conjugates that do not bind in the test area (due to not being bound by the anti-human hemoglobin) are captured by an immobilized anti-immunoglobulin antibody located in the control area designated C, resulting in the formation of a pink line. Because the mobile antibody-dye complex is normally at a molar excess compared to the human hemoglobin, the control line is expected to appear and indicates proper performance of the test device in the absence of a pink band in the test area. REAGENTS, SUPPLIES, & MATERIALS 1. ABAcard HemaTrace Test Kit containing devices and extraction buffer (Abacus Diagnostics, Catalog #708424) WORKSHEETS 1. ABAcard HemaTrace Worksheet SAMPLE REQUIREMENTS Samples will consist of portions of fabric, swabs, and other materials that may contain human blood-based stains. Typically, the need for HemaTrace testing by this procedure will be indicated by previous presumptive chemical testing. Indications other than presumptive chemical testing may apply in specific case circumstances. PROCEDURE NOTE: Before beginning this procedure, assure that all critical reagents, solutions and materials have been QCed and have not passed their expiration date. NOTE: Test results obtained with the HemaTrace test devices must be observed and confirmed by a second qualified analyst. The second analyst will record the verification of results by writing their initials and the date of testing on the ABAcard HemaTrace worksheet.
Serology Procedures Manual OneStep ABAcard HemaTrace Test for the Forensic Identification of Human Blood (2.4.2009)
A. Sample Extraction 1. Place the sample in the provided vial of extraction buffer. a. For swabs, the amount of swab used is up to the discretion of the analyst. i. For a swab that gives a strong presumptive reaction, or less of a swab may be sufficient. ii. For a swab that gives a weaker presumptive reaction, up to of a swab may be sufficient. iii. The maximum amount that can be used for this test is of a swab, so as to preserve a sufficient amount of the sample for DNA testing. When multiple swabs are available for testing, more than one swab may be tested. The maximum amount that can be used from any of the available swabs is of a swab. b. For stains on fabric, use approximately 0.5 cm2. i. This amount may be adjusted based upon the intensity of the presumptive reaction and the size of the stain. 2. Fully saturate and submerge the substrate in the extraction buffer by centrifuging and/or vortexing. 3. Incubate at room temperature for at least 5 minutes. . Older bloodstains can be extracted for an extended period of time at the discretion of the analyst. The extraction period may be extended up to 24 hours in length. a. The test may be performed immediately after extraction or stored at 4oC for up to one week or at -20oC indefinitely. B. ABAcard HemaTrace Test Procedure 1. The HemaTrace test is performed in a batch-wise manner. 2. Each batch includes the following samples processed in parallel: a. Test Sample Extracts: Test sample extracts are prepared as described in Part A. 150 l of each extract is used in the HemaTrace test. b. Positive Control: The positive control is 150 l of HemaTrace Blood Standard 1:1,000,000 dilution of whole blood. The extraction buffer used as a negative control must be the same lot of extraction buffer used to extract the test samples. The positive control may be stored at 4oC for up to one week or stored indefinitely at 20oC in 200 l aliquots. c. Negative Control: The negative control is 150 l of extraction buffer. The extraction buffer used as a negative control must be the same lot of extraction buffer used to extract the test samples. 3. Allow the sample to warm to room temperature if it has been refrigerated or frozen. 4. Prepare an ABAcard HemaTrace Worksheet for recording the results of the test. 5. Label the devices with the sample identifier. a. For test sample extracts, this will be the FL # and sample #. b. For controls, this will be POS (or ) or NEG (or ). 6. Initial and date each test device. 7. Vortex the test sample and positive control extracts. 8. Transfer 150 l of the positive control (HemaTrace blood standard, 1:1,000,000 dilution of whole blood) into the sample well of the corresponding test device ( control).
9. Transfer 150 l of the sample extract into the sample well of the corresponding test device. a. If several extracts are being processed in a batch-wise fashion, apply the extracts to the corresponding devices in quick succession avoiding delays. 10. Transfer 150 l of the negative control (extraction buffer) into the sample well of the corresponding test device ( control). 11. Incubate the test devices for 10 minutes at room temperature. 12. At the end of the 10 minute incubation, read the results of the tests by visual examination of the HemaTrace test devices and record them on the result sheet (for further discussion of the interpretation and responses to test results, see Section C). NOTE: Abnormal or unusual test results that fall outside the following guidelines will be brought to the attention of a supervisor for evaluation on a case-by-case basis. a. Positive result. If there are 2 pink lines, one in the test area T and one in the control area C, then the test is positive. b. Weak positive result. If there are 2 pink lines, one in the test area T and one in the control area C, and the T line is fainter than the T line on the positive control test, then the test is weak positive. c. Negative result. If there is a pink line in the control area C, but no line in the test area T, then the test is negative. d. Inconclusive. If there is no pink line in the control area C, then the test is inconclusive. 13. Immediately after reading the results, photograph the devices to document the test. a. NOTE: The digital photograph serves only to document the test. The test results are based upon direct visual examination of the test devices, not the photographs. Faint lines may not be visible in the hardcopy photographs. 14. Print the photograph of the devices and attach it to the worksheet(s). C. Evaluation of Test Results and Responses to Test Results 1. Evaluate the negative control. a. The negative control is expected to give a negative result. b. If the negative control fails (i.e., gives a positive or inconclusive result), then the test is unreliable and must be repeated. 2. Evaluate the positive control. a. The positive control is expected to give a positive result. b. If the positive control fails (i.e., gives a negative result or inconclusive result), then the test is unreliable and must be repeated. 3. Evaluate the result for the test samples. a. Positive result. i. Per the manufacturer, a positive result indicates that the human hemoglobin level is above 0.5 g/ml in the extract. ii. A positive result is reported as Positive. b. Weak positive result. i. A weak positive result may not be readily visible when the device is photographed. ii. A weak positive result is reported out as Positive. c. Negative result. i. A negative result may indicate that:
a. Human hemoglobin is not present in the extract b. Human hemoglobin is present at a level below the detection limit of the assay c. Human hemoglobin is present at an extremely high level, resulting in the High Dose Hook Effect ii. If the analyst determines that the extraction was effective and that the negative result was due to the High Dose Hook Effect, he/she may dilute the sample 1:10 (20 l sample extract, 180 l extraction buffer) and repeat the test. a. If the diluted sample gives a negative result, and if there is insufficient sample quantity to re-extract the sample, then the test result is Negative. b. If the result is positive, then the test result is Positive. c. If the diluted sample gives a negative result, and if there is sufficient sample quantity to re-extract the sample, then extract more of the sample and repeat assay. i. If the result is positive, then the test result is Positive. ii. If the result is again negative, then the test result is Negative. d. Inconclusive result. i. If the test is inconclusive (due to the absence of a pink band in the Control area), and there is sufficient sample to repeat the test, then the test must be repeated. ii. If the test is inconclusive, and there is insufficient sample to repeat the test, then the test result is Inconclusive. D. Documentation of Test Results 1. The ABAcard HemaTrace Worksheet is available as an electronic (.doc) form. a. Header and sample information may be filled in electronically. The form will then be printed. b. Alternatively, the header and sample information may be filled in manually on a hardcopy printout of the form. 2. Test results are to be filled in manually on a hardcopy form. a. If the batch includes test samples from multiple cases, then one case packet will contain the original worksheet. The remaining case packets will contain photocopies of the worksheet. 3. Photographs of HemaTrace test devices are to be printed out and attached to the HemaTrace worksheet. a. If there are test samples from multiple cases in the batch, then print sufficient photographs for each report packet. b. Original photographs (not copies) must be included in each report packet. E. Reporting Results and Conclusions 1. Description of results. Written descriptions of the test results should clearly indicate whether human hemoglobin was observed or not observed. 2. Conclusions. As appropriate, statements of conclusions should indicate the interpretation of the test results as opposed to the results themselves. These conclusions will correspond to the expert opinion that will be offered in the case of testimony.
SPECIES OF ORIGIN DETERMINATION USING THE OUCHTERLONY DOUBLE DIFFUSION METHOD (2.4.2009) PRINCIPLE Antigen and antibody reactants are allowed to diffuse toward each other through a gel medium to produce visible precipitin arcs which permits antigen identification. I. Preparation of stain extracts A. Apparent bloodstained cuttings of minimum size (approximately 1 mm x 1 mm) are extracted in 10-30 l of Millipore water or 5% ammonium hydroxide (in the case of 5% ammonium hydroxide, a 5% NH 4 OH extraction blank will then need to be run as a negative control). 1. Only stains testing presumptive positive for blood should be tested for species determination. 2. Cutting size and water volume may be adjusted slightly, if necessary, to account for stain intensity and size. 3. Extraction should have a straw color. 4. Extraction time will depend on the condition and concentration of the stain. Most stains will take no longer than 1 hour to extract. If the extraction is performed overnight, the extraction should be stored at 4oC. 5. Older or degraded stains may be extracted at 37oC. Procedure A. Gels are prepared for origin determination by punching wells using a template. 1. For Petri dish testing, a rosette pattern will be employed using the center well for antibody and the six surrounding wells for samples and controls. 2. Wells should be about 5 mm or less in diameter and approximately 5 mm apart. B. Example sample placement. 1. Apply samples in the following pattern: Antiserum center circle Positive control 1 and 4 Unknown sample or negative control 2, 3, 5, and 6 1 6 5 4 2 3
II.
Serology Procedures Manual SPECIES OF ORIGIN DETERMINATION USING THE OUCHTERLONY DOUBLE DIFFUSION METHOD (2.4.2009)
2. Two positive controls (for human species test - 1:1000 dilution of whole blood) must be present in each rosette. 3. At least one negative control (extracting medium) must be present on each plate. 4. This procedure may be used to determine the origin of many species provided the proper anti-sera can be located, a positive control is available and run with testing to be conducted, and the antiserum is checked for quality in the same manner as the human antiserum. C. Place approximately 5-7 l (until the well is full but not overflowing) of the stain extracts, antiserum, and controls in the appropriate wells of the gel. D. Place in a 37oC incubator for 4 to 8 hours, at room temperature overnight, or refrigerate overnight (may be left refrigerated over a weekend). E. Results are read by viewing the gels with oblique transmitted light, or gel may be stained by placing stain directly over gel until lines of identity are stained, then pouring remainder of stain off gel. 1. Staining optional. Stain in 0.1% Coomassie Blue stain for 15-30 minutes. Destain until the background is clear. III. Interpretation of Results A. Positive results occur as precipitin arcs between antiserum and stain extract (or known serum) wells. 1. All positive controls must test positive. 2. All negative controls must not contain a precipitin arc. 3. If positive controls are blank or negative controls contain a precipitin, then the test should be repeated on a new gel plate. 4. Two analysts must evaluate the plate and initial and date the worksheet. 5. If results are negative, a second test should be performed on the same or a new extract to confirm the negative result. B. Record the results as positive or negative on the Species worksheet. 1. Include the worksheet in the case packet. 2. If the batch includes test samples from multiple cases, then one case packet will contain the original worksheet. The remaining case packets will contain photocopies of the worksheet. 3. Results of testing should be included in the Results section of the SA kit and/or Examination worksheet.
Serology Procedures Manual SPECIES OF ORIGIN DETERMINATION USING THE OUCHTERLONY DOUBLE DIFFUSION METHOD (2.4.2009)
BUCCAL SWAB STORAGE PROCEDURES (2.4.2009) PRINCIPLE Buccal swabs are dried and stored frozen as standards for future DNA analysis. Only open one buccal swab collection kit at a time. I. Procedures for SWIFS buccal swab collection kits A. Check out buccal swab kits from Evidence Registration. B. Remove buccal swab collection kit from mailing packaging, if necessary. C. The collection kit envelope should be sealed. Note on the buccal swab worksheet if the kit is sealed or unsealed. D. Cut open the buccal swab collection kit envelope without breaking the evidence seal. E. Remove the buccal swab collection kit card which contains the swab envelope. F. Label all envelopes and the collection kit card with the laboratory number, item number, your initials, and the date of processing. G. Complete the case number information on the worksheet using information from the submittal sheet. Record from whom and when the evidence was received on the worksheet. H. The signature from the subject information section should be used as the identification of the person from whom the sample was collected. If the signature is illegible, the printed portion of the subject information should be used. If neither of these is available, the name printed on the swab envelope can be used. I. Note the number of swabs on the worksheet. J. Place swab(s) in a ziploc bag labeled with the laboratory number and item number. Place this bag in a heat seal bag labeled with the laboratory number, item number, complainants name, your initials, the date, and the subjects name if different than the complainant. Seal the bag and initial and date your seal. K. Make photocopies of the buccal swab collection kit card and swab envelope. L. Store the swab(s) in the -20oC freezer. M. Seal the packaging materials, placing initials and date on the evidence seal. Return the packaging material to Evidence Registration.
II. Procedures for receiving buccal swabs not in standard buccal swab collection kits A. Make copies of packaging material that can be used to identify swab(s). B. Use packaging material or information on Submittal Sheet to identify owner of the swab(s). Use the case number and item number if a name is not given. C. Follow the above listed steps.
Serology Procedures Manual BUCCAL SWAB STORAGE PROCEDURES (2.4.2009)
SEXUAL ASSAULT CLOTHING EVIDENCE ANALYSIS (2.4.2009)

PRINCIPLE
Large items of evidence may be screened for the presence of biological fluids such as seminal fluid or blood using a series of presumptive tests. Initial rapid screening of large items may be performed by visual inspection using visible light, magnification, and/or using an alternate light source (ALS). Specific areas or stains may be further analyzed using chemical tests. Areas testing presumptive positive may then be retained and preserved for further testing, which may include confirmatory testing and/or DNA analysis.
WORKSHEETS
1. Evidence examination worksheet

SAMPLE REQUIREMENTS
The following procedures will be followed for examination of sexual assault clothing and other evidence relating to sexual assault cases. However, specific case circumstances that may require deviation from these procedures will be allowed under analyst discretion. Any trace evidence associated with the item(s) will be handled according to the procedures for the collection, documentation, and preservation for trace evidence. If apparent blood stains are observed on the evidence item, follow the procedure outlined in the LMG Test for Blood.
PROCEDURE
A. Evidence Examination Worksheet 1. Fill out an evidence examination worksheet, including all submittal and reagent QC information at the top. a. If items are submitted by different persons, on different days, or under different tags, this information must be clearly indicated on the worksheet. b. If items are examined on different days, this must be clearly indicated on the worksheet, along with the reagent QC information for each day. c. If two different lots of reagent were used to screen the item(s), then both reagent lots must be clearly indicated on the worksheet. d. If test areas are removed from the same item on different days, this must be clearly indicated on the worksheet. e. If more than one analyst examines an item, the examination worksheet must be cosigned by each analyst performing the examination. 2. For each item of evidence, include the item #, a description of the packaging and any other appropriate and relevant information. a. Any information written or stamped on the evidence or packaging regarding the location of collection, agency numbers when appropriate, date of collection, or other information of importance should be recorded on the worksheet. b. If the examination is not performed in the Forensic Biology Laboratory (e.g. the Firearms Unit), this information must be noted on the worksheet. B. Examination of the Evidence
Serology Procedures Manual Sexual Assault Clothing Evidence Analysis (2.4.2009)
1. Examine clothing or other items using the alternate light source (ALS). a. Follow the procedure for examination with the ALS. NOTE: If the entire item will be tested using the Brentamine Fast Blue B test for ACP, then the item need not be screened with the ALS. b. Areas identified with the ALS will be sampled by swab analysis and analyzed for ACP using the Brentamine Fast Blue B test. i. If no ALS positive areas are identified, then the entire item must be tested for ACP. ii. Items that have a high level of background fluorescence must be completely tested for ACP. 2. Examine the evidence for ACP. Follow the procedure for testing using the Brentamine Fast Blue B test. All of the following will be tested: a. ALS positive areas i. If ALS positive areas are identified on the item, the areas are prominently visible above any background fluorescence, and at least one ALS positive area is identified as ACP positive, then the remainder of the item need not be tested for ACP. b. Visible stains c. Areas of critical importance such as the crotch of pants or areas indicated during communication with investigators d. The remainder of the item when necessary (see above B.1.b.i. and B.1.b.ii.). 3. If apparent blood stains are observed on the item, perform the LMG test on the stain using the procedure outlined in the LMG Test for Blood. NOTE: The LMG Test for Blood may be performed on the same swab after the Brentamine test. 4. Retain test areas as necessary for additional analysis. a. Test areas may be removed and stored for possible DNA analysis. Follow the procedure for Collection and Storage of Test Samples. b. In the event that an ambiguous result is obtained with the ACP test (e.g. very weak positive, or color change immediately after 15 sec), it is at the analysts discretion if additional testing is necessary. The analyst may choose to: i. Extract a portion of the stain and look for the presence of spermatozoa. Stains should be extracted according to the procedure outlined in the protocol, OneStep ABAcard p30 test procedure. ii. Perform confirmatory testing on the stain following the OneStep ABAcard p30 test procedure. c. When an apparent blood stain is identified on the item (i.e., stain gives a positive result for the LMG Test for Blood), a small portion of an apparent blood stain may be removed in order to perform a confirmatory test for blood (Ouchterlony Species Test or OneStep ABAcard HemaTrace Test). 4. Record results on the worksheet. See Guidelines for Description and Results sections below (C and D). 5. Place the FL#, item number, date of analysis, and initials of examiner performing analysis on the garment. a. When possible, place the information near the tag of an item for easy identification. (e.g., shirts - inside back under collar, pants/panties - inside back under waistline, etc.) i. Avoid placing identification information in a place where it may be removed from the item. ii. If the information is written in an unusual place, document on the Evidence
Examination worksheet where the FL# is written. iii. If the information can not be written on the item, document on the Evidence Examination worksheet that the FL# is not written on the item. C. Guidelines for Description section of the Evidence examination worksheet 1. Items should be described in the following order: a. A description of the packaging, including any relevant information written or printed on the package (see above A.2.a.) b. A listing of the item using the format, item#: item (e.g. 1:Panties). c. A physical description of the item which should include but not be limited to the following information: color and style of item and information provided on the manufacturers label (written in quotations), including brand, size and type of fabric d. Presentation and general appearance of the item e. Visible stains f. Defects g. Debris/hairs associated with the item 2. Abbreviations may be included in the Description section. A list of acceptable abbreviations is given in Appendix 2. D. Guidelines for Results section of the Evidence examination worksheet 1. The Results section should be spaced apart from the Description section on the worksheet. 2. The Results section should clearly indicate the results, positive or negative, of any tests performed. i. Positive areas retained for further analysis should be clearly described in the Results section and designated, T 1 , T 2 , etc., and enclosed in a box, or otherwise set apart from the text. ii. Additional comments describing the test area (e.g. visible stain, 2 swabs, etc...) may be written next to the box. 3. Abbreviations may be included in the Results section. A list of acceptable abbreviations is given in Appendix 2. 4. In some cases, a diagram of the item should be included in the Results section of the worksheet. Diagrams are helpful when an item contains more than one stain, type of stain, multiple analyses have been performed, or multiple test areas have been retained. Diagrams should include but not be limited to the following information: . A brief description of what is being depicted, the viewpoint of the diagram (i.e.outside/front view) i. The location of areas testing positive (these areas will be color coded using green only for ACP positive areas, red only for CAT positive visible stains, and yellow only for ALS positive areas) . Noteworthy stains, writings, defects (including tears, bullet holes, manufacturer defects, etc...) . The location of the FL#, if the FL# is written in an unusual place 5. Test areas should be clearly marked on the diagram in the manner described previously. The test area sampled should be marked on the diagram and a notation should be made as to the manner of sampling (i.e. sample or cutting). 6. A photograph of the item may be included instead of a diagram. This may be particularly
useful in cases where complex stains are observed, or multiple viewpoints need to be shown. i. The photograph may be labeled in the same manner described for a diagram, with all test areas marked appropriately. ii. Photographs may be standard or digital. iii.Photographs should include a ruler in order to establish scale.
BLOOD EVIDENCE ANALYSIS (2.4.2009) PRINCIPLE Evidentiary items can be screened for blood by performing a presumptive chemical test using the leucomalachite green (LMG) reagent. A color change is observed when LMG is oxidized in the presence of hemoglobin. The presence of human blood is confirmed by using the ABAcard HemaTrace Test for Blood or the Species of Origin Test using the Ouchterlony Double Diffusion Method. Samples of appropriate areas are collected and stored for further analysis. I. Evidence Examination Worksheet A. Fill out an evidence examination worksheet, including all submittal and reagent QC information at the top. i. If items are submitted by different persons, on different days, or under different tags, this information must be clearly indicated on the worksheet. ii. If items are examined on different days, this must be clearly indicated on the worksheet, along with the reagent QC information for each day. iii. If two different reagents were used to analyze an item, QC results for both reagents must be clearly indicated on the worksheet. iv. If test areas are removed from the same item on different days, this must be clearly indicated on the worksheet. v. If more than one analyst examines an item, the examination worksheet must be cosigned by each analyst performing the examination. B. For each item of evidence, include the item #, a description of the packaging and any other appropriate information. i. Any information written or stamped on the evidence or packaging regarding the location of collection, agency numbers when appropriate, date of collection when appropriate, or other information of importance should be recorded on the worksheet. ii. If the location of examination is not the Forensic Biology Laboratory (e.g. Firearms and Toolmarks Unit) this information must be noted on the worksheet. II. Evidence Examination A. Determine if multiple analyses are required before proceeding (e.g. blood spatter analysis and blood) and coordinate examination with the analyst from the appropriate laboratory. B. QC LMG reagent with a positive (human blood diluted 1:10,000) and negative control swab before use. C. Examine visible stains first. Use magnification, as necessary, to visualize any stains. i. Sample all visible stains by swab analysis method. ii. Perform the LMG presumptive test for blood. iii. Analyze the remainder of the item using the LMG test (see Collection and Storage of Test areas, for retaining positive stains). iv. If it is necessary to preserve a portion of an item for other analyses (e.g. Latent Prints), those portions need not be tested unless deemed necessary after those
Serology Procedures Manual BLOOD EVIDENCE ANALYSIS (2.4.2009)
analyses are complete. This information must be noted on the worksheet (e.g. This portion preserved for possible latent print analysis per A.J. Jumper 9-11-01). D. Record results on the worksheet. See Guidelines for Descriptions and Results sections under Sexual Assault Clothing Evidence Analysis. E. Place the FL#, item number, date of analysis, and initials of examiner performing analysis on the item when possible. If not possible, note on the Evidence Examination worksheet. F. If there is sufficient sample quantity, retain a small portion of appropriate test areas for confirmatory testing. i. Perform either the ABAcard HemaTrace Test for Blood or the Species of Origin Test using the Ouchterlony Double Diffusion Method. ii. Record the results on the worksheet next to the test area that was analyzed. iii. Date and initial the results.
Serology Procedures Manual BLOOD EVIDENCE ANALYSIS (2.4.2009)
EVIDENCE HANDLING AND ANALYSIS PROCEDURES FOR SEXUAL ACITIVITY KITS (2.4.2009) The following procedures will be implemented regarding the collection and storage of Sexual Activity kits (SA kits) and processing of the evidence with the kits by the Forensic Biology Unit (FBU). I. Collection of Sexual Activity Kits from Parkland Memorial Hospital (PMH) Police Locked Cabinet A. Internal Chain. After retrieving the SA kits from the PMH Police lock box, the internal chain of custody will be initiated by the analyst or authorized representative of Physical Evidence. 1. The internal chain will reflect the cabinet the SA kit was retrieved from and be signed by the person that retrieved it. 2. Any clothing retrieved from the locked box will require a separate internal chain and be submitted to the Evidence Registrar. 3. No internal chain should be initiated for kits which contain only a doctors report and no evidence. B. Submittal sheets and chains of custody. A submittal sheet should accompany each SA kit and any clothing retrieved from the locked box. 1. If a submittal sheet is not found within the kit or with the clothing, one should be initiated by the analyst or authorized representative of Physical Evidence. 2. The submittal form will be removed from the kit. 3. The submittal form should be stamped with the time and date of receipt. 4. Write on the submittal form whether or not the seals on the SA kit and clothing are intact. 5. The original chain of custody and DPD tag should stay with the SA kit and/or clothing. C. The doctors report. The doctors report will be removed from the kit and photocopied for the case file. 1. The FL # should be indicated on both the originals and photocopies (may be written on original before copying). 2. Write the word clothing on the top front page of the doctors report if any clothing has been submitted. 3. Write the word Tox on the top front page of the doctors report if a Toxicology kit has been submitted. 4. Return the original doctors report to the manila folder in the hallway labeled Attn: G. Ackerman, OB/GYN Dept,, UTHSCD or SWIFS Childrens Medical Center or another appropriate envelope. The original doctors report from male SA kits will be retained in the case file. 5. If a doctors report only and no other evidence is received, retain a copy of the doctors report in the case file. a. Use the appropriate report format to notify the investigating agency. Any paperwork submitted with the doctors report will remain in the case file. The

EVIDENCE HANDLING AND ANALYSIS PROCEDURES FOR SEXUAL ACITIVITY KITS (2.4.2009)
D.
E.
F.
G.
doctors report will be assigned an FL #, but no item #. No internal chain should be initiated. Sexual Activity Kit Log. A SA kit log should be filled out each time kits are retrieved. 1. Fill out the date on the top portion of the sexual activity log and fill in complainants name, agency service #, and submitting agency in applicable areas. In the case of Dallas Police Department cases, include the tag number for each sexual activity kit submitted by Dallas P.D. 2. All the SA kits submitted to Evidence Registration at a given time should be listed on the log sheet. 3. Indicate in the appropriate area whether or not clothing has been submitted with the kit. 4. Indicate in the appropriate area the date of offense. 5. Notate on the log sheet those SA kits containing an oral rinse. 6. Notate on the log sheet male SA kits. 7. Write the agency service # on all SA kits. 8. Write the Dallas P.D. tag # on all SA kits submitted by Dallas P.D. 9. Give the completed SA kit log sheet to the Evidence Registrar. The Evidence Registrar will assign the FL # and the beginning item # for each SA kit and clothing and return the completed log sheet to the analyst. 10. The evidence listed on each SA kit log sheet should be entered in the FBU lab computer log book as soon as possible. The original SA kit log sheet will be filed in the appropriate notebook within the FBU laboratory. Toxicology Kit Log. A toxicology kit log should be filled out each time toxicology kits are retrieved. 1. Fill out the date on the top portion of the sexual activity log and fill in the complainants name, agency service #, and submitting agency in applicable areas. In the case of Dallas Police Department cases, include the tag number for each toxicology kit submitted by Dallas P.D. 2. Be sure that all edges of the toxicology kit are sealed. If not, seal all the edges. Initial and date the new seals. 3. Write the agency service # on all toxicology kits. 4. Write the Dallas P.D. tag # on all toxicology kits submitted by Dallas P.D. Transfer of the SA kit to Evidence Registration. After the necessary paperwork has been removed from the SA kit, the kit will be resealed by the custodian of the kit and will be transferred to Evidence Registration. In the event that the kits can not be immediately transferred to Evidence Registration, the SA kits may be transferred to another analyst or temporarily stored in a communal area such as the community locked cabinet and/or evidence storage refrigerator located within the FBU. Note: SA kits containing an oral rinse should be stored in the FBU evidence storage refrigerator. Transfer of the Toxicology kit to Evidence Registration. Transfer the toxicology kits to the Evidence Registrar as soon as possible. In the event that the kits can not be immediately transferred to Evidence Registration, the toxicology kits may be transferred to another analyst or temporarily stored in a communal area such as the secured refrigerator within the FBU.

II. Temporary Storage of Sexual Activity Kits within the Forensic Biology Laboratory SA kits may be temporarily stored prior to and during analysis within designated communal storage areas within the FBU. A. The SA kits must be sealed while in communal storage areas. B. SA kits may be stored in communal storage areas. These areas will be located within the laboratory and will be accessible only to Forensic Biology personnel. C. Alternatively, SA kits may be stored in the analyst's personal locked storage cabinet (kits need not be resealed when locked up under the analyst's personal control). III. Transfer of SA kit components to other units A. Internal chain. When possible, the original internal chain of custody should remain with the SA kit. However, at times it may be necessary to transfer components of the kit to other laboratories within the Institute. 1. The original internal chain may be transferred with the components (e.g. blood samples to Toxicology) and a copy retained with the kit until the components are returned. 2. Transfers may be documented with original signatures on a copy of the internal chain 3. Alternatively, a new internal chain may be initiated by the analyst in order to transfer components of the kit when necessary. Every effort should be made by the analyst to minimize the number of internal chains. B. SA kit. Items may be removed from the SA kit for analysis at a later time (e.g. smears) or analysis by another unit (e.g. hair evidence). 1. When the analyses are complete, the items will be returned to the kit. IV. Guidelines for screening SA kits. There are two general types of SA kits received into the FBU for analysis, kits containing an oral rinse and kits lacking an oral rinse. In cases that involve an alleged oral assault, the oral rinse may contain DNA from the assailant in the form of sperm cells. SA kits containing an oral rinse should be refrigerated until the evidence sample can be stabilized. V. Inventory of SA kits The contents of each SA kit should be inventoried and item numbers assigned to the evidence, unless the kit is to be returned the investigating agency without analysis. A. Start a SA kit worksheet. Fill out all submittal and complainant information. B. Write FL#, item #, date, and initials on each item. Number the items following the order listed on the worksheet if possible.

C. If "vaginal", "anal", "oral", or other, is not indicated on a swab box or other item, then the item should be listed simply as "swab", or appropriate name, on the worksheet.
VI.
Analysis of SA kits The FBU has a 30 day policy for the analyzing of evidence contained within a SA kit. No analysis will be performed on items with the kit unless a request is received from the investigating agency within 30 days of the submittal of the kit. After 30 days, if no request for analysis has been received, the SA kit and contents, and all other evidence associated with the case that was submitted to the FBU, will be returned to the investigating agency with no analysis. Agencies that may not be familiar with the FBUs 30 day policy will be contacted at the end of the 30 day period to determine if the agency requests analysis of the kit. Analyze all swabs and smears using appropriate testing methods described in the FBU Serology Procedures Manual. Individual case circumstances may necessitate a deviation from the standard kit processing guidelines, and analyst discretion should be used. All items in the kit are potential biohazards and should be analyzed using the precautions for blood-borne pathogens. Items submitted will be analyzed according to the following guidelines: A. Swabs. All swabs will be analyzed for the presence of acid phosphatase (ACP) using the Brentamine Fast Blue B test, unless submitted as a standard (e.g. saliva sample) or if other information specific to the case indicates that no analysis is necessary. 1. If a positive ACP or p30 result is obtained with the swabs, or the corresponding smear is positive for spermatozoa, store the swabs in a ziploc baggie labeled with the unique FL #, and item #. Place the baggie in a plastic envelope labeled with the FL #, item #s, complainant's name, analyst's initials, and storage date. Heat seal the envelope. Date and initial the seal. File the envelopes in the evidence storage freezer. Follow proper procedure concerning documentation of evidence transfer on the internal chain. Note the results of the analysis, stored date, and initials on the SA kit worksheet. 2. If a swab is submitted for ACP analysis without a corresponding smear, a smear should be prepared from the swab or other means should be used to confirm the presence of semen. 3. If a swab tests positive for ACP and the corresponding smear is negative for spermatozoa, then the other smear (if a pair is received) must also be stained and examined. If both smears are negative, then a portion of the swab should be eluted and examined according to the procedures outlined for the ABAcard p30 test. 4. If the doctor's report indicates Doctor saw sperm, document the name of the physician who observed spermatozoa on the worksheet. Analyze the vaginal swabs and smears. Store the swabs regardless of results and note stored on worksheet. If swabs and smears are negative, then a portion of the swab may be

eluted and examined according to the procedures outlined for the ABAcard p30 test. 5. If a swab appears to contain blood, note this on the SA kit worksheet. Depending on case circumstances (e.g., the evidence was collected from a child), the swab should be tested for blood and stored even if the ACP test is negative. Confirmatory testing may be performed on a portion of the swab if the sample quantity is sufficient. B. Smears. One of each pair of vaginal, anal, oral, and/or oral rinse smears will be stained using the Christmas tree staining method and microscopically examined for the presence of spermatozoa. 1. If a single smear is received, it will be stained and analyzed. The oral smear need not be examined unless specific case circumstances dictate. The oral smear must be examined if the kit is an autopsy kit, or if no oral rinse has been submitted. 2. If circumstances dictate (e.g. scant cells on the smear), the other smear may be stained and analyzed. 3. If a swab tests positive for ACP and the corresponding smear is negative for spermatozoa, then the other smear (if a pair is received) must also be stained and examined. 4. If both an oral smear and oral rinse are submitted, the oral rinse should be analyzed according to the following: a. Centrifuge the sample for 2-3 minutes or adequate time to form pellet. b. Transfer pellet onto a clean glass slide using a clean pipette. c. Label the slide with the FL#, item# and last name, first initial of the complainant. d. Allow the sample to dry. e. Stain and examine as described above. f. The remaining pellet should be transferred onto a sterile cotton swab, and the swab air dried. This swab will remain in the SA kit unless testing indicates that the swab should be stored for possible further analysis (see below). g. In the event that spermatozoa are observed on the smear prepared from the oral rinse, or the oral swab tests positive for the presence of ACP, the swab containing the remainder of the oral rinse pellet should be stored as described for swabs (VI.A.1.). C. Toxicology. When blood and/or urine samples are submitted in the SA kit, those samples should be transferred to the Toxicology Unit (Tox) in the event that the doctor's report indicates that toxicological analysis is requested, the investigating or submitting agency requests analysis, or other information available to the analyst suggests that analysis should be performed. 1. Transfer one 10 ml red top urine and two 10 ml grey top blood tubes in a plastic biohazard ziploc bag sealed with evidence tape. 2. Label the biohazard bag with the FL# and item #. 3. If no grey top tubes are submitted, a purple top tube may be sent to toxicology instead. If this is the case, store a sample of the blood on cloth before submitting it to Toxicology. 4. Fill out the Tox request information in the comments section of the internal chain, which includes a description of the items, the number of tubes submitted, and Serology Procedures Manual
which analyses are requested. (e.g. 2-10 ml grey blood, 1-10 ml red urine; D/A, GHB/ROHYP (circle one or both analyses). 5. Have the Toxicology Evidence Registrar (or a toxicologist) sign the internal chain. Leave an internal chain and samples with the Evidence Registrar. a. Return all samples to the SA kit after Tox analyses are complete. 6. If Tox is not requested, then seal each blood and urine tube with masking tape to prevent leakage (some urine tubes come already taped and need not be re-taped). Place all tubes in a plastic biohazard bag and return to the SA kit. D. Blood standard storage. This procedure should be performed according to the guidelines concerning exposure to blood-borne pathogens. In the event that a positive test for blood or seminal fluid is obtained on appropriate items, or in response to a request for DNA analysis, the blood standard from the complainant should be stored. 1. Store the complainant blood card following the procedure outlined in the Collection and Storage of Test Areas. The blood card standard must be stored separately from any other items collected and/or retained. 2. If the complainant blood standard consists of a tube of blood, blood should be placed on cloth prior to storage. a. Using a sterile pipette, transfer ~0.3 - 0.5 mL blood from purple top (or other when appropriate) tube on a square of sterile cloth. Allow to dry thoroughly. b. Store in a labeled, sealed baggie in the FB freezer following the procedure for swab storage (VI.A.1.). c. Check the box, "On cloth", on the SA worksheet, or fill out the Blood Standard Storage form for out of county kits. List the item #, date and initial in the "stored" area at the bottom of the SA kit worksheet. d. Document on the internal chain transfer of evidence to the FB freezer for storage. e. The report will state that "a portion of item ..." has been retained. If the report has already been issued before the standard has been stored, a new report need not be issued for storage of the blood standard only. E. Hair evidence. Pubic hair combings will be opened and examined for hair and debris. 1. Note on the SA kit worksheet if hair/debris was observed by checking the appropriate box, " yes, no, other". 2. Remaining hair samples will be labeled and returned to the kit. This also includes autopsy cases. 3. Each report with hair items will include statements concerning analysis and disposition which are appropriate according to the current Trace evidence handling protocols (see Examination and Processing of Hair Combings in Sexual Assault Casework). F. Debris Collection. Debris collection envelopes must be opened and the contents inventoried. 1. Analyze the contents when appropriate. 2. The contents of the envelope should be described on the SA worksheet, and listed in the report.

G.
H.
I.
J.
K.
3. Apparent hair or debris should be documented according to the policies concerning Trace Evidence collection (see Examination and Processing of Hair Combings in Sexual Assault Casework and Collection of Trace Evidence). Autopsy kits. For analysis of autopsy kits, look at the submittal form to check if kit is to be stored or analyzed. If analysis is requested, analyze all swabs and smears following the procedures described above. Suspect kits. Suspect kits containing items collected from the suspect may be submitted to the FB Laboratory. 1. Fill out a Suspect kit worksheet, including all submittal and inventory information. 2. Store blood on cloth the same as for SA kits using the Blood Standard Storage form. 3. Store but do not analyze penile swabs, oral swabs, or saliva samples. Out of county kits. SA kits may be submitted from agencies outside of Dallas County and usually are from a different manufacturer and/or style than the sexual activity kits issued by S.W.I.F.S. 1. Fill out the standard SA kit worksheet noting the manufacturer of the kit at the top. 2. Place the complainant's blood on cloth using the Blood Standard Storage form. Condoms. Condoms may be submitted inside an SA kit or as additional items with the case. Condoms should be analyzed carefully because of their fragile nature. 1. Condoms may have complainant epithelial cells on one side (the outside) and suspect epithelial cells and/or seminal fluid on the opposite side. Therefore, sampling should be done on each side separately. 2. Examination with an Alternate Light Source may be performed in order to determine which side contains seminal fluid. Clothing. Often items of clothing, usually collected from the complainant, or other items of evidence are submitted with a SA kit. These items need not be examined at the time of kit examination. 1. In the event that a positive test result for seminal fluid is observed for items in the SA kit, any clothing or other items accompanying the kit may be returned to the proper agency without analysis. 2. The clothing should be retained if it is possible that multiple assailants are involved, if the complainant has had consensual intercourse prior to the assault, or if other circumstances dictate that analysis of the clothing is necessary. 3. If the clothing is returned without analysis, then the following statement (or equivalent) should be included in the report indicating the reason: No serological or hair/fiber analysis was performed on item * due to the detection of spermatozoa and/or acid phosphatase on items obtained from the sexual assault kit. 4. If the clothing will be examined, procedures for sexual assault clothing examination, should be followed.

EXAMINATION AND PROCESSING OF HAIR COMBINGS IN SEXUAL ASSAULT CASEWORK (2.4.2009) PRINCIPLE This document describes the procedures used by the Forensic Biology Unit (FBU) in regard to combings submitted with sexual activity kits (non-autopsy) submitted in investigations of sexual assaults. NOTE: The procedures described are to be applied to standard routine casework. These procedures may be modified by the examiner, based upon individual circumstances and/or specific instructions of the investigator.
SAMPLE A sexual activity kit (non-autopsy) submitted for examination, that includes hair combing evidence.
DEFINITIONS For the purpose of these procedures, the following definitions will be used. 1. Trace evidence. Particulate and/or fibrous material that is loosely associated with an evidence item, and which by its nature may be easily dislodged from an item, or transferred from one item to another item. 2. Debris. Undifferentiated trace material, either adhering to an item or dislodged from an item. 3. Loose debris. Undifferentiated trace evidence that becomes dislodged from an item during routine processing procedures. 4. Adhering debris. Undifferentiated particulate or fibrous material that is attached to an item. 5. Apparent hair. Material that in the opinion of the examiner has a gross visual (nonmicroscopic) appearance consistent with a human or animal hair, and that does not have the appearance consistent with a fiber (natural or synthetic). 6. Apparent fiber. Material that in the opinion of the examiner has the gross visual (nonmicroscopic) appearance consistent with a synthetic or natural fiber, and that does not have the appearance consistent with a hair (human or animal).
Serology Procedures Manual EXAMINATION AND PROCESSING OF HAIR COMBINGS IN SEXUAL ASSAULT CASEWORK (2.4.2009)
7. Apparent hair or fiber. Material that in the opinion of the examiner has a gross visual (non-microscopic) appearance consistent with either hair (human or animal) or fiber (natural or synthetic).
I. PROCEDURES A. Examination of hair combing evidence Following inventory of the evidence containers submitted in the kit, the examiner will open containers labeled as hair combing evidence, and perform a preliminary evaluation of their contents for the presence of hairs and/or fibers or other debris. 1. The combing evidence will be examined visually without the aid of a microscope or other magnifying device. The contents of the container will be categorized as follows: a. Debris. The examiner observes undifferentiated trace material, but no material whose appearance is consistent with being hairs or fibers. Apparent hairs. The examiner observes material whose appearance is consistent with a human or animal hair, and is not consistent with a fiber (natural or synthetic). Apparent fibers. The examiner observes material whose appearance is consistent with a fiber (natural or synthetic), and is not consistent with a human or animal hair.
2.
b.
c.
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Following examination, the evidence will be returned to the container, and the container will be secured. a. The container will be closed, sealed with evidence tape, and initialed by the examiner. The container will be labeled with the laboratory case number, the item number, the examiners initials, and the date.
b.
II. BENCH NOTES 1. The kit worksheet prepared during evidence examination will note the presence of any obvious debris in the combings. The kit worksheet will note if the hair evidence was transferred to the TEU for examination. 2
2.
III. REPORTING 1. When apparent hair, fiber or other debris is noted in the hair combings, the serology report will include in the results section an appropriate statement. E.g., a. Debris was observed in item 4. The possible evidentiary value of this material is unknown. In the event that analysis is required, please contact the Trace Evidence Unit. Apparent hair was observed in item 4, and items 4 and 5 have been transferred to the Trace Evidence Unit.
b.
2.
The disposition of hair evidence will be noted in the section of the report: Disposition of Evidence.
COLLECTION, PRESERVATION, AND DOCUMENTATION OF TRACE EVIDENCE (2.4.2009) PRINCIPLE This document describes the policies and procedures that will be followed by the Forensic Biology Unit (FBU) in regard to trace evidence that may be present on items of evidence submitted for the purpose of routine screening or blood and other body fluid stains. DEFINITIONS For the purpose of these procedures, the following definitions will be used. 1. Trace evidence. Particulate and/or fibrous material that is loosely associated with an evidence item, and which by its nature may be easily dislodged from an item, or transferred from one item to another item. 2. Debris. Undifferentiated trace material, either adhering to an item or dislodged from an item. 3. Loose debris. Undifferentiated trace evidence that becomes dislodged from an item during routine processing procedures. 4. Adhering debris. Undifferentiated particulate or fibrous material that is attached to an item. 5. Apparent hair. Material that in the opinion of the examiner has a gross visual (nonmicroscopic) appearance consistent with a human or animal hair, and that does not have the appearance consistent with a fiber (natural or synthetic). 6. Apparent fiber. Material that in the opinion of the examiner has the gross visual (nonmicroscopic) appearance consistent with a synthetic or natural fiber, and that does not have the appearance consistent with a hair (human or animal). 7. Apparent hair or fiber. Material that in the opinion of the examiner has a gross visual (non-microscopic) appearance consistent with either hair (human or animal) or fiber (natural or synthetic). POLICIES 1. Inasmuch as items of evidence submitted to the FBU for the identification of body fluids may contain trace evidence of possible evidentiary value, all evidence examination procedures used by the FBU will be performed in a manner that avoids the loss of or contamination of such material. 2. Analysis of trace evidence including the evaluation of its evidentiary value and identification of its component materials (e.g., human and animal hair, synthetic and natural fibers) is
Serology Procedures Manual COLLECTION, PRESERVATION, AND DOCUMENTATION OF TRACE EVIDENCE (2.4.2009)
performed by the Trace Evidence Unit (TEU), and all questions regarding the analysis and evidentiary value of trace evidence will be referred to the staff of the TEU. 3. Evidence examinations will be performed in a manner that avoids contamination of the trace evidence present on an item. a. A fresh piece of examination paper will be used for each evidence item examined. b. A new pair of examination gloves will be used for each evidence item examined. c. The bar used for hanging large evidence items will be cleaned with 10% bleach before each use, and between each evidence item examined. I. Procedure for Securing trace evidence 1. Following screening of an evidence item, any loose debris will be secured in one of the following ways: a. Paper bindle method i. Loose debris will be collected into a sheet of paper, or a portion of a sheet of paper, triple-folded in two directions to form a pocket. ii. The bindle will be placed in one of the following containers: (1) coin envelope (2) other envelope (3) zip lock bag (4) heat-sealable bag iii. The debris container will be closed and sealed as appropriate. iv. The debris container will be sealed with evidence tape, and the evidence tape will be initialed by the examiner. b. Post-it note method i. Small amounts of loose debris will be collected on the adhesive strip of a post-it note sheet. ii. The post-it note will be folded in-half upon the adhesive strip, thereby immobilizing the debris. iii. The post-it note will be placed in one of the following containers: a. coin envelope b. other envelope c. zip lock bag d. heat-sealable bag iv. The debris container will be closed and sealed as appropriate. v. The debris container will be sealed with evidence tape, and the evidence tape will be initialed by the examiner. 2. The debris container will be labeled with the following information: FL#, item #, complainant name, initials, date of collection. 3. The debris container will be sealed with evidence tape and initialed by the examiner. 4. The debris container will be returned with the evidence item to its original container. II. Bench notes
1. The bench notes prepared during evidence examination will note the following regarding the presence of trace evidence. a. The presence of any obvious adhering debris on the item, described as follows: i. debris ii. apparent hair iii. apparent fiber iv. apparent hair and/or fiber b. The collection and securing of any loose debris from the item, described as follows: i. debris ii. apparent hair iii. apparent fiber iv. apparent hair and/or fiber 2. When loose debris is collected, a notation will be made in the bench notes. E.g., Debris: Packaged, returned with item III. Evidence release & chain of custody When the evidence is released to the evidence registration unit or to another laboratory unit, the chain of custody document will note that a package of debris is included with the evidence item. IV. Reporting 1. When loose debris is collected from an item, or when significant adhering debris is noted on an item, the serology report will include in the results section an appropriate statement. E.g., a. Debris, including apparent hairs, was collected from item 2. The possible evidentiary value of this material is unknown. In the event that analysis is required, please contact the Trace Evidence Unit. b. Debris, including apparent hairs, was noted on the following items: 2, 4, 6. The possible evidentiary value of this material is unknown. In the event that analysis is required, please contact the Trace Evidence Unit. 2. The disposition of collected debris evidence will be noted in the section of the report: Disposition of Evidence. E.g., a. Debris collected from item 2 will be returned to the investigating agency with that item. b. The following items and associated debris will be returned to the investigating agency: 2, 7.
STORAGE OF EVIDENCE IN THE FORENSIC BIOLOGY FREEZER (2.4.2009) PRINCIPLE The following procedures will be implemented regarding the storage of evidence in the Forensic Biology Unit -20C secured freezers (FB freezers). I. Transfer of evidence by Forensic Biology personnel. A. The internal chain of custody will reflect transfer of evidence collected by Forensic Biologists during analysis to the -20C freezer within the Forensic Biology Laboratory. This policy also includes evidence submitted to the Laboratory for storage until further analysis (e.g. buccal swabs). B. The internal chain will also reflect removal of evidence from the FB freezer for further analysis or transfer/release. i. Every effort should be made to record all evidence transfers on a single original internal chain of custody. ii. If this is not possible, a new internal chain may be initiated for specific evidentiary items when necessary. Every effort should be made to minimize the number of internal chains in the case file. iii. All evidence transfers must be documented in accordance with the Institutes Evidence Handling procedures policy. iv. FB freezers located outside of the FBU will be locked with access to keys restricted to Forensic Biology personnel. II. Transfer of evidence collected by others units to the FB freezer. Biological evidence collected by examiners from other units within the Physical Evidence Section may be transferred to the FB freezer for storage. These evidentiary must be accompanied by an internal chain of custody and transferred to a member of the Forensic Biology Laboratory who will then transfer them to the FB freezer. The chain will then be filed in the appropriate case file.
Serology Procedures Manual STORAGE OF EVIDENCE IN THE FORENSIC BIOLOGY FREEZER (2.4.2009)
APPENDIX 1. Reagents and Solutions: Preparation, Storage, and QC (2.4.2009) Anti-human Antisera Purchased as lyophilized powder from Seri Reconstitute as described by Seri Add 1 ml of Millipore or sterile water Freeze in aliquots at -20oC QC Procedure: 1. Run a species plate according to the Ouchterlony procedure. 2. Include 2 wells of human blood standard and 1 well of water for positive and negative controls, respectively. 3. Both positive controls must test positive and the negative control must be blank. 4. Each new vial of reconstituted antisera will be tested against the host animal before put into use. 5. Record QC date and initials on the antisera storage container. Brentamine Reagent Materials Anhydrous Sodium Acetate Glacial Acetic Acid Deionized Water Sodium Alpha-Naphthyl Acid Phosphate (Sigma N-7000) o-Dianisidine, Tetrazotized; Fast Blue B (Sigma D-9805) Brentamine Buffer 1. Dissolve 6 g of Anhydrous Sodium Acetate in 500 ml of Deionized Water. 2. Add 2 ml of Glacial Acetic Acid to the solution. 3. Check that the pH of the solution is 5.0 0.2. This is the optimum pH for the catalytic activity of the ACP enzyme. 4. Store in the refrigerator in a capped bottle with the preparation date until ready to use. Brentamine Reagent 1. In a clean glass amber bottle, combine the following: 0.01 g 0.002 g Sodium Alpha-Naphthyl Acid Phosphate o-Dianisidine, Tetrazotized
2. Add 10 ml of the Brentamine Buffer to the bottle and mix until powder is dissolved.
Serology Procedures Manual APPENDIX 1. Reagents and Solutions: Preparation, Storage, and QC (2.4.2009)
Brentamine Reagent QC Brentamine reagent should be prepared fresh daily. Each preparation of the Brentamine Reagent must undergo quality control (QC) before initial use. This is performed by adding a drop of the reagent to a positive control swab containing semen at a 1:1,000 dilution and observing a color change within 15 seconds. A negative control swab moistened with distilled water should be included in the QC test. No color change should be observed on this swab. Christmas Tree Stain Materials Deionized Water Aluminum Sulfate Nuclear Fast Red Picric Acid Solution Indigo Carmine Dye Nuclear Fast Red Solution (NF) 1. 2. 3. 4. 5. Dissolve 15 g of Aluminum Sulfate in 600 ml of Deionized Water. Heat on a stir plate, as necessary, to dissolve. Immediately add 0.3 g of Nuclear Fast Red to the solution. Cool to room temperature and filter through Whatman paper. Place solution into an amber colored stoppered bottle.
This solution lasts approximately 3-6 months stored at room temperature away from sunlight. Prepare fresh, as necessary. Picroindigocarmine Solution (PICS) 1. Dissolve 2 g of Indigo Carmine Dye into 600 ml of Picric Acid solution. 2. Stir until dissolved. 3. Place the solution into an amber colored stoppered bottle. This solution lasts approximately 4 months stored at room temperature away from sunlight. Prepare fresh, as necessary. Coomassie Blue Staining Solution 0.1% Coomassie Blue (R250) dissolved in destain solution Destain Solution 20 ml Ethanol
20 ml Deionized Water 4 ml Acetic Acid Leuchomalachite Green Reagent Materials Leucomalachite Green (p, p1 benzylidinebis-N, N1 dimethylaniline) (Sigma #L-7257) Sodium Perborate (J.T. Baker #3811-05) Sodium Bisulfite (J.T. Baker #3556) Acetic Acid (reagent grade) Deionized Water 40% Acetic Acid Solution Combine 100 ml of Acetic Acid with 150 ml of Deionized Water. This can be pre-made in larger quantities for ongoing usage, and stored in a labeled container with a lid. Sodium Bisulfite Solution Dissolve 1.48 g of Sodium Bisulfite in 8 ml of 40% Acetic Acid solution. LMG Reagent Solution 1. Prepare a flask, wrap with foil if flask is to be left in the light for a lengthy period of time. 2. Dissolve 0.54 g of Leucomalachite Green into 112 ml of 40% Acetic Acid solution. 3. Mix well by placing on a stirring plate with a stir bar in solution. 4. Titrate the Sodium Bisulfite solution into the LMG reagent solution by adding drops slowly. Note: This is done over a period of time, occasionally adding drops at intervals of approximately one minute apart until all of the Sodium Bisulfite solution is added. 5. Place the reagent solution in (2) amber bottles and secure the tops. Properly label, date and initial the bottles and store in the refrigerator. Note: One batch will make two bottles. Also, place a white mark on the tops of the lid bulbs for easy identification of reagent solution at the bench. LMG Activator Solution 1. Dissolve 6 g of Sodium Perborate into 112 ml of 40% Acetic Acid solution. 2. Place the activator solution in (2) amber bottles and secure the tops. Properly label, date and initial the bottles and store in the refrigerator.
LMG QC 1. LMG reagent and activator solutions are QCd in pairs labeled with the lot number and designated pair A, B, etc. QC is performed by adding a drop of reagent solution, immediately followed by a drop of activator solution to a positive control swab prepared from a 1:10,000 dilution of human blood. A blue-green color change should be observed within 5 seconds. If no color change is observed, the reagent pair does not pass QC. Determine whether the failure is due to expiration of the control swab before preparing new reagent. 2. Test a negative control swab with each reagent pair, as described above. The negative control swab is a sterile cotton swab moistened with deionized water. No color change should be observed after addition of either LMG reagent or activator solution. If a color change is observed, the reagent pair does not pass QC. 3. Each batch of LMG reagent and activator solutions should be QCd after preparation. Record QC results in the Forensic Biology Unit QC logbook. Additionally, each reagent pair must be QCd immediately prior to use. 4. Over time, the efficacy of the LMG reagent will begin to deteriorate. This deterioration will be evident as the reagent will begin to turn blue in color. At this point, the reagent must not be used any longer. Approximate shelf life of the LMG solutions is one month. p30 Semen Standard Purchased as a lyophilized powder from Seri Reconstitute as directed: Add 0.5 ml of sterile water Freeze at -20oC QC using same procedure as for p30 Test Reagents Species Gel Buffer 1. Dissolve 0.533 g of Diethylbarbital in 100 ml hot water 2. Add 3.5 g Sodium Diethylbarbital, 0.512 g Calcium Lactate and q.s. to 500 ml. Adjust pH to 8.6. 3. Store refrigerated.
Species Gel Plates 1. 2. 3. 4. Dissolve 1.4 g of Agar Noble agar in 70 ml of water in a flask and bring to a boil. Add 70 ml of gel buffer mix by rotating the flask. Pipette 7 ml of this mixture into the bottom of a Petri dish. Allow plates to cool and refrigerate in a moisture box at 4oC. 4
This preparation will make about 20 plates in Petri dishes. Species Gel Plate Stain/Destain 1. Dissolve 0.1 g Coomassie blue in 100 ml of destain solution (outlined below). 2. Destain solution: 20 ml Methanol (50 vols.) 20 ml Distilled Water (50 vols.) 4 ml Acetic Acid (10 vols.)
QC TEST: P30 TEST REAGENTS (other than kits) 1. Onestep ABAcard p30 Test devices. Purchase from Abacus Diagnostics. Store at 4oC. QC test prior to use in casework. 2. Deionized water. Use Millipore water or equivalent. 3. Semen/p30 standard. The semen/p30 standard may be crude semen containing p30 (SERI p30 electrophoresis standard), purified p30 (Sigma Chemical), or equivalent. The standard should be resuspended in water (if necessary), and diluted with water to a final concentration of 4 ng/ml based on nominal concentrations provided by the manufacturer. Diluted standard will be aliquoted and stored at -20oC. Prior to use in casework, each batch of the standard should be QC tested using non-expired p30 test devices.
QC TEST: ABACARD P30 KITS PRINCIPLE This document describes the quality control procedure for the OneStep ABAcard p30 Test Kits for the Identification of Semen. This procedure is to be followed each time a new lot of test kits is received into the laboratory. REAGENTS, SUPPLIES, MATERIALS 1. Seminal Fluid Standard 1:25,000 (prepared from SERI R563 Semen Standard) 2. ABAcard p30 Test Kit (Abacus Diagnostics p/n 308332) 3. Deionized water WORKSHEETS 1. ABAcard p30 worksheet SAMPLE REQUIREMENTS Positive control The positive control is 200 l of Semen Standard diluted to a final concentration of 4 ng/ml (1:25,000). The positive control will be stored in aliquots of approximately 80 l at -20oC. Negative control The negative control is 200 l of deionized water. PROCEDURE NOTE: Each new lot of kits must pass QC prior to use in the testing of evidence samples for the presence of seminal fluid. NOTE: The manufacturers specified expiration date of test kits may be extended 30 days based upon successful QC testing. Re-certification of kits may be repeated two additional times (90 days total). A. Log in of Kits 1. Each new lot of kits is given a laboratory specific tracking number. a. Enter the new lot of kits into Forensic Biology QC notebook. b. The laboratory specific lot number will be recorded on the p30 Worksheet each time the p30 test is performed on samples. B. ABAcard p30 Test QC Procedure 1. Allow the samples to warm to room temperature if they have been refrigerated or frozen. 2. Prepare an ABAcard p30 Worksheet for recording the results of the test. 3. Label the test devices with the sample identifier. This will be POS (or ) for the positive control and NEG (or ) for the negative control.
4. Initial and date each test device. 5. Vortex the positive control extract. 6. Transfer 200 l of the positive control (Seminal Fluid Standard - 1:25,000) to the corresponding test device ( control). 7. Transfer 200 l of the negative control (Deionized Water) to the corresponding test device ( control). 9. Incubate the test devices for 10 minutes at room temperature. 10. At the end of the 10 minute incubation, read the results of the tests by visual examination of the p30 test devices and record them on the results sheet (for further discussion of the interpretation and responses to test results, see Section C). NOTE: Abnormal or unusual test results that fall outside the following guidelines will be brought to the attention of a supervisor for evaluation on a case-by-case basis. a. Positive result. If there are 2 pink lines, one in the test area T and one in the control area C, then the test is positive. b. Weak positive result. If there are 2 pink lines, one in the test area T and one in the control area C, and the T line is fainter than the T line on the positive control test, then the test is weak positive. c. Negative result. If there is a pink line in the control area C, but no line in the test area T, then the test is negative. d. Inconclusive. If there is no pink line in the control area C, then the test is inconclusive. 11. Immediately after reading the results, photograph the devices to document the test. a. NOTE: The digital photograph serves only to document the test. The test results are based upon direct visual examination of the test devices, not the photographs. Faint lines may not be visible in the hardcopy photographs. 12. Print the picture of the test devices and attach it to the worksheet. C. Evaluation of Test Results and Responses to Test Results 1. Evaluate the negative control. a. The negative control is expected to give a negative result. b. If the negative control fails (i.e., gives a positive result), then the test must be repeated. i. If the negative control gives a negative result, the lot passes the positive QC. ii. If the negative control fails (i.e., gives a positive result), then the failure must be reported to a supervisor for evaluation. 2. Evaluate the positive control. a. The positive control is expected to give a positive result. b. If the positive control fails (i.e., gives a negative result or inconclusive result), then the test must be repeated. i. If the positive control gives a positive result, the lot passes the positive QC. ii. If the positive control fails (i.e., gives a negative result or inconclusive result), then the failure must be reported to a supervisor for evaluation. D. Documentation of QC Test Results 1. The ABAcard p30 Worksheet is available as an electronic (.doc) form.
2. 3. 4. 5.
a. Header and sample information may be filled in electronically. The form will then be printed. b. Alternatively, the header and sample information may be filled in manually on a hardcopy printout of the form. Test results are to be filled in manually on a hardcopy form. Photographs of p30 test devices are to be printed out and attached to the p30 worksheet. Test results are to be written in the Forensic Biology Unit QC logbook. File the worksheet in the Forensic Biology Unit QC notebook.
QC TEST: ABACARD HEMATRACE KIT

PRINCIPLE
This document describes the quality control procedure for the OneStep ABAcard HemaTrace Test Kits for the Identification of Human Blood. This procedure is to be followed each time a new lot number of test kits is received into the laboratory.
REAGENTS, SUPPLIES, & MATERIALS
1. ABAcard HemaTrace Test Kit containing devices and extraction buffer (Abacus Diagnostics, Catalog #708424) 2. Deionized Water
WORKSHEETS
1. ABAcard HemaTrace worksheet

SAMPLE REQUIREMENTS
Positive Control: The positive control is 150 l of HemaTrace Blood Standard 1:1,000,000 dilution of whole blood. The positive control may be stored at 4oC for up to one week or stored indefinitely at -20oC in 200 l aliquots. Negative control The negative control is 150 l of the extraction buffer provided with the new lot of kits.
PROCEDURE
NOTE: Each new lot of kits must pass QC prior to use in the testing of evidence samples for the presence of human blood. NOTE: The manufacturers specified expiration date of test kits may be extended 30 days based upon successful QC testing. Re-certification of kits may be repeated two additional times (90 days total). C. Log in of Kits 1. Each new lot of kits is given a laboratory specific tracking number. a. Enter the new lot of kits into Forensic Biology QC notebook. b. The laboratory specific lot number will be recorded on the HemaTrace Worksheet each time the HemaTrace test is performed on samples. D. ABAcard HemaTrace Test QC Procedure 1. Allow the samples to warm to room temperature if they have been refrigerated or frozen. 2. Prepare an ABAcard HemaTrace Worksheet for recording the results of the test.
10
3. Label the test devices with the sample identifier. This will be POS (or ) for the positive control and NEG (or ) for the negative control. 4. Initial and date each test device. 5. Vortex the positive control extract. 6. Transfer 150 l of the positive control (HemaTrace blood standard, 1:1,000,000 dilution of whole blood) to the corresponding test device ( control). 7. Transfer 150 l of the negative control (extraction buffer) to the corresponding test device ( control). 9. Incubate the test devices for 10 minutes at room temperature. 10. At the end of the 10 minute incubation, read the results of the tests by visual examination of the HemaTrace test devices and record them on the result sheet (for further discussion of the interpretation and responses to test results, see Section C). NOTE: Abnormal or unusual test results that fall outside the following guidelines will be brought to the attention of a supervisor for evaluation on a case-by-case basis. a. Positive result. If there are 2 pink lines, one in the test area T and one in the control area C, then the test is positive. b. Weak positive result. If there are 2 pink lines, one in the test area T and one in the control area C, and the T line is fainter than the T line on the positive control test, then the test is weak positive. c. Negative result. If there is a pink line in the control area C, but no line in the test area T, then the test is negative. d. Inconclusive. If there is no pink line in the control area C, then the test is inconclusive. 11. Immediately after reading the results, photograph the devices to document the test. a. NOTE: The digital photograph serves only to document the test. The test results are based upon direct visual examination of the test devices, not the photographs. Faint lines may not be visible in the hardcopy photographs. 12. Print the picture of the test devices and attach it to the worksheet. C. Evaluation of Test Results and Responses to Test Results 1. Evaluate the negative control. a. The negative control is expected to give a negative result. b. If the negative control fails (i.e., gives a positive or inconclusive result), it may indicate that: i. The extraction buffer provided with the kits contains human hemoglobin at concentrations above 0.5 g/ml. ii. The lot of test devices is faulty. c. If the negative control gives a positive result, then repeat the test using two different negative controls. i. Negative control #1: 150 l of the extraction buffer provided with the new lot of kits. ii. Negative control #2: 150 l of deionized water. d. If both negative controls give a negative result, the lot passes negative QC. e. If either negative control #1 or negative control #2 fails (i.e., gives a positive or inconclusive result), then the failure will be reported to a supervisor for evaluation. 2. Evaluate the positive control.
11
a. The positive control is expected to give a positive result. b. If the positive control fails (i.e., gives a negative result or inconclusive result), then the test must be repeated. i. If the positive control gives a positive result, the lot passes the positive QC. ii. If the positive control fails (i.e., gives a negative result or inconclusive result), then the failure must be reported to a supervisor for evaluation. D. Documentation of QC Test Results 2. The ABAcard HemaTrace Worksheet is available as an electronic (.doc) form. a. Header and sample information may be filled in electronically. The form will then be printed. b. Alternatively, the header and sample information may be filled in manually on a hardcopy printout of the form. 2. Test results are to be filled in manually on a hardcopy form. 3. Photographs of HemaTrace test devices are to be printed out and attached to the HemaTrace worksheet. 4. Test results are to be written in the Forensic Biology Unit QC logbook. 5. File the worksheet in the Forensic Biology Unit QC notebook.
12
APPENDIX 2. ABBREVIATIONS FOR BENCH NOTES (2.4.2009)

The following are acceptable abbreviations to use on the worksheet: ACP CBB HFB BPB CAT NA NFA NR Omni PSA/P30 QNSFAS Quant WPB WK T# k PMH OB/GYN CMC SA FL# QC SWIFS Occ. Mod. LMG - Acid Phosphatase - Cardboard box - High fluorescent background - Brown paper bag - Catalytic, peroxidase-like activity, denotes LMG test result - No analysis - No further analysis - No results - Omnichrome or other alternate light source - Prostate specific antigen - Quantity Not Sufficient for Further Analysis Serology - Acid phosphatase quantitation - White paper bag - Weak - Test area and number - Negative result - Positive test result - Parkland Memorial Hospital - Obstetrics/Gynecology - Childrens Medical center - Sexual Assault - Forensic Laboratory Number - Quality Control - Southwestern Institute of Forensic Sciences - Occasional - Moderate - Leucomalachite Green - Pass - Fail
Serology Procedures Manual APPENDIX 2. ABBREVIATIONS FOR BENCH NOTES (2.4.2009)
APPENDIX 4. QUALITY CONTROL FOR SEROLOGICAL EXAMINATION AND SAMPLE PROCESSING (2.4.2009) PRINCIPLE The following are quality control measures that will be followed during examination of evidence samples in order to minimize contact with blood-born pathogens and to prevent crosscontamination of evidentiary items. These procedures conform to the guidelines established in the Institute Biological Exposure Plan, the Institute Quality Management Program, and the Forensic Biology Quality Management Program. I. Protective Wear A. All analysts must wear proper laboratory attire appropriate for the analyses they are performing. i. Lab coats must be worn for all analyses. ii. Disposable gloves must be worn for all analyses and changed as frequently as is necessary. B. Safety glasses and other safety equipment such as face masks must be worn when appropriate. II. Examination Materials A. 10% bleach must be prepared fresh daily B. All swabs used for evidence examination and collection must either be purchased sterile and the package unopened prior to use, or sterilized by autoclaving prior to use. C. Scalpel blades must be purchased sterile and the packaging unopened prior to use. D. All water used for sample elution must be autoclaved or Millipore. Deionized water may be used for moistening of swabs prior to swab analysis. III. Examination Areas A. Examination areas will be cleaned with 10% bleach before and after each case, and as necessary in between items. i. The bench-top on which the analysis is to be performed will be wiped down with 10% bleach before beginning work on evidence from each case. ii. The bench-top may be cleaned with 10% bleach between examinations of items within a case as deemed necessary by the analyst (e.g. extremely dirty or bloody items, or items seized at different locations). iii. After analysis on items from a case is complete, the bench-top will be cleaned with 10% bleach. B. Clean examination paper will be placed under each item of evidence examined. C. Examination of evidence will precede one item at a time.
Serology Procedures Manual APPENDIX 4. QUALITY CONTROL FOR SEROLOGICAL EXAMINATION AND SAMPLE PROCESSING (2.4.2009)
i.
ii.
Only a single item of evidence will be removed from the packaging at the same time and be placed in the same work area for examination Multiple samples retained from an item or various items must not be open at the same time.
IV.
Removal of Test Areas A. All utensils including scissors and forceps used to remove positive test areas must be cleaned with 10% bleach before and after each use B. Standards must not be stored in the same plastic envelope as questioned samples.
V.
Disposal of Materials A. All evidence submitted to the FB Laboratory is a potential biohazard and should be examined according to the guidelines listed in the Biological Exposure Manual. All materials coming in contact with evidence (e.g. swabs, gloves, etc.) must be disposed of in a Biohazard bag or waste container. B. Sharp items must be disposed of in a sharps container or glass disposal container as appropriate. C. Chemicals should be disposed of according to the Institute Hazardous Chemical policies.
Serology Procedures Manual APPENDIX 4. QUALITY CONTROL FOR SEROLOGICAL EXAMINATION AND SAMPLE PROCESSING (2.4.2009)
SOUTHWESTERN INSTITUTE OF FORENSIC SCIENCES DALLAS, TEXAS
BUCCAL SWAB KIT

TAG#__________________ FL# __________________________ DCME# __________________________
AGENCY CASE# __________________________ COMPLAINANT NAME __________________________
RECEIVED FROM _____________________________ ON ________________________ .
KIT SEALED:
YES
NO
KIT SPECIMEN RECEIVED:
___________ BUCCAL SWAB STANDARD (
COMPLAINANT SUSPECT OTHER _____________________________ _____________________________
STORED BY: _____________________________ ON _____________________________ .
REV. 06-2001
SOUTHWESTERN INSTITUTE OF FORENSIC SCIENCES Dallas, Texas

Brentamine Lot # _______ POS: __ Pass NEG: __ Pass Date_______ Initials______ FL ________________________ LMG Lot # ________ POS: __ Pass NEG: __ Pass Date_______ Initials_____ Comp ______________________
Agency _____________________ Received from ________________________ on _________________. Tag # ______________ Agency # ______________________ Offense _____________________
Examiner ______________________________
Stored: _______________________
_________________________ Date ______________________________
SOUTHWESTERN INSTITUTE OF FORENSIC SCIENCES Dallas, Texas

FL # ___________________________ Comp ___________________________
Examiner ______________________________
Stored: _______________________
_________________________ Date ______________________________
Sexual Activity Kit Log

Date and Time: ___________________ Complainant Name / Date of Offense Agency Name / Service #
Laboratory #
Evidence Retrieved SA Kit Tox Kit Oral Rinse SA Kit Tox Kit Oral Rinse SA Kit Tox Kit Oral Rinse SA Kit Tox Kit Oral Rinse SA Kit Tox Kit Oral Rinse SA Kit Tox Kit Oral Rinse SA Kit Tox Kit Oral Rinse SA Kit Tox Kit Oral Rinse yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no yes no
Page ____ of ____
SOUTHWESTERN INSTITUTE OF FORENSIC SCIENCES SEXUAL ASSAULT WORKSHEET TAG #: ______________ Brentamine Reagent Lot #: _________________ QC pos: neg: Initials _____ Date _________ FL #: ____________________ Agency: _____________________ Agency #: ________________________ Offense: ________________________ Complainant Name:_________________________ Recd From/By: Age/Race/Gender: ____ / ____ / ____
PMH Police lock box / ________ Date and Time:________________ _______________________________ Kit Sealed: Yes No Seal Initialed: Yes No SEXUAL ASSAULT KIT SPECIMENS RECEIVED: From Doctors Report:__________________, observed sperm. : Underwear : Vaginal swab(s) : Vaginal smear(s) : Anal swab(s) : Anal smear(s) : Oral swab(s) : Oral smear(s) : Oral rinse ( ( ( ( ( ( ( ) ) ) ) ) ) ) : Urine sample ___________ : Blood sample ___________ On cloth : Blood card : Pubic hair combings Yes No Other : Pubic hair standard : Head hair combings Yes No Other : Head hair standard : Debris collection Microscopic Examination __________________________________ __________________________________ __________________________________
Acid Phosphatase Screen __________________________________ __________________________________ __________________________________
NOTES: ____ : Oral rinse - This item was centrifuged. From the resulting pellet, the following were made: ____ Slide(s), ____ Swab(s). These remain in kit unless otherwise noted.
Stored/Date: ________________ ________________ ________________
Analyst ______________________________ Date ______________________________
Toxicology Collection Kit Pick-Up Log

Laboratory # Complainant Name Agency Name / Service # Retrieved from PMH Police Lock box Other PMH Police Lock box Other PMH Police Lock box Other PMH Police Lock box Other PMH Police Lock box Other PMH Police Lock box Other PMH Police Lock box Other PMH Police Lock box Other Above listed Toxicology kits: Retrieved by ____________________________ on ________________________. Transferred to ___________________________ on ________________________. Transferred to ___________________________ on ________________________. Page ____ of ____

SWIFS FBU Serology Procedures Manual v2.0 (02.04.2009) - Re Formatted

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Southwestern Institute of Forensic Sciences Criminal Investigation Laboratory

Forensic Biology Unit Serology Procedures Manual, Version 2.0 (2.4.2009)

Serology Procedures Manual (2.4.2009)

Serology Procedures Manual (2.4.2009) Table of Contents

1/16/2003 7/3/2003 1/24/2008

Sliter Sliter Sliter

Serology Procedures Manual Corrections & Revisions Log (2.4.2009)

Serology Procedures Manual Corrections & Revisions Log (2.4.2009)

Serology Procedures Manual BACKGROUND ON SEROLOGICAL TESTING OF FORENSIC EVIDENCE (2.4.2009)

ACP x 4 * ACP x 4 * ACP wk x 2*

Serology Procedures Manual MICROSCOPIC EXAMINATION OF SMEARS FOR SPERMATOZOA (2.4.2009)

Coordinates 35.9 x 100.1

Initials and Date KRD 5/9/08 KRD 5/9/08

Serology Procedures Manual MICROSCOPIC EXAMINATION OF SMEARS FOR SPERMATOZOA (2.4.2009)

Serology Procedures Manual COLLECTION AND STORAGE OF TEST AREAS (2.4.2009)

Serology Procedures Manual COLLECTION AND STORAGE OF TEST AREAS (2.4.2009)

Serology Procedures Manual

Serology Procedures Manual BUCCAL SWAB STORAGE PROCEDURES (2.4.2009)

SEXUAL ASSAULT CLOTHING EVIDENCE ANALYSIS (2.4.2009)

1. Evidence examination worksheet

Serology Procedures Manual Sexual Assault Clothing Evidence Analysis (2.4.2009)

Serology Procedures Manual BLOOD EVIDENCE ANALYSIS (2.4.2009)

Serology Procedures Manual BLOOD EVIDENCE ANALYSIS (2.4.2009)

Serology Procedures Manual

Serology Procedures Manual

Serology Procedures Manual

Serology Procedures Manual

Serology Procedures Manual

Serology Procedures Manual

QC TEST: ABACARD HEMATRACE KIT

1. ABAcard HemaTrace worksheet

APPENDIX 2. ABBREVIATIONS FOR BENCH NOTES (2.4.2009)

Serology Procedures Manual APPENDIX 2. ABBREVIATIONS FOR BENCH NOTES (2.4.2009)

SOUTHWESTERN INSTITUTE OF FORENSIC SCIENCES DALLAS, TEXAS

BUCCAL SWAB KIT

AGENCY CASE# __________________________ COMPLAINANT NAME __________________________

RECEIVED FROM _____________________________ ON ________________________ .

KIT SPECIMEN RECEIVED:

___________ BUCCAL SWAB STANDARD (

COMPLAINANT SUSPECT OTHER _____________________________ _____________________________

STORED BY: _____________________________ ON _____________________________ .

SOUTHWESTERN INSTITUTE OF FORENSIC SCIENCES Dallas, Texas

SOUTHWESTERN INSTITUTE OF FORENSIC SCIENCES Dallas, Texas

Sexual Activity Kit Log

Page ____ of ____

Acid Phosphatase Screen __________________________________ __________________________________ __________________________________

Stored/Date: ________________ ________________ ________________

Analyst ______________________________ Date ______________________________

Toxicology Collection Kit Pick-Up Log

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AGENCY CASE# COMPLAINANT NAME

RECEIVED FROM _____ ON .

COMPLAINANT SUSPECT OTHER _ _

STORED BY: _ ON _ .

Page of

Acid Phosphatase Screen __

Stored/Date:

Analyst Date