SPECIMEN COLLECTION AND TRANSPORT

MEDICAL LABORATORY FACULTY MEDICINE BRAWIJAYA UNIVERSITY

MICROBE ATTACHMENT (ADHERENCE) COLONIZATION INVASION / NO PRODUCE METABOLITE PRODUCTS/ NO INFECTION (DISEASE)

ROLE OF CLINICAL MICROBIOLOGIST AND CLINICIAN

CLINICAL MICROBIOLOGIST
COLABORATION/ SERVICES REQUIRED

CLINICIAN

INFECTIOUS DISEASES

SERVICES REQUIRED BY CLINICIANS FROM CLINICAL MICROBIOLOGIST

1. RELEVANT INFORMATION NEEDED TO MAKE MEDICAL DECISIONS 2. ESTABLISHMENT OF GUIDELINES FOR PROFER

SPECIMENS COLLECTION
3. ENSURING TIMELY SPECIMENS TRANSPORT 4. IDENTIFICATION AND SUSCEPTIBILITY TESTING OF MICROBE 5. RAPID REPORTING OF TESTS RESULT 6. CLOSE CONSULTATION

SPECIMEN COLLECTION AND TRANSPORT

1. CRITICAL CONSIDERATION, BECAUSE THE QUALITY OF A LABORATORY WORKS MAY BE LIMITED BY THE NATURE OF A SPECIMENS AND CONDITION 2. SPECIMEN SHOULD BE OBTAINED TO PRECLUDE (menghindari) OR MINIMIZE THE POSSIBILITY OF INTRODUCING EXTRANEOUS MICROORGANISMS 3. CAREFUL SKIN PREPARATION BEFORE PROCEDURE SUCH AS BLOOD CULTURE AND SPINAL TAPS, FOR MAJOR APPROACHES ARE USED TO RESOLVE THIS PROBLEM: - CLEAN SKIN SURFACE WITH GERMICIDE USING ENOUGH FRICTION - STRART CENTRALLY AND GO OUT IN EVER ENLARGING CIRCLE - REPEAT THIS SEVERAL TIME USING A NEW SWAB EACH TIME - ALKOHOL 70% IS SATISFACTORY FOR SKIN , BUT A FULL 2 MINUTES OF WET CONTACT IS NEDEED.

IODINE (2% TINCTURE OF IODINE AND POVIDONE-IODINE MORE QUIKCLY (1 MINUTE) AND ARE EFFECTIVE AGAINST SPOREFORMING ORGANISMS

SPECIMEN COLLECTION AND TRANSPORT

4. WHENEVER POSSIBLE, SPECIMENS SHOULD BE OBTAINED BEFORE ANTIMICROBIAL AGENTS HAVE BEEN ADMINISTRED 5. SUSPECTED ORGANISMS IS MOST LIKELY TOBE FOUND, WITH A LITTLE EXTERNAL CONTAMINATION AS POSIBLE 6. PERSONEL MUST BE INSTRUCTED TO LABEL CAREFULLY AND SPECIMEN SUBMITTED TO LABORATORY, INCLUDING DATA: PATIENTS NAME,

HOSPITAL NUMBER, DATE AND TIME COLLECTION, ANTIBIOTIC TREATMENT, AND THE EXACT NATURE OF SPECIMEN

Suggested Transport Media General Comments

Medium Stuarts Medium Amies Medium Utility Most aerobic and some facultative anaerobes. Most anaerobic and facultative anaerobes GC General purpose medium for transport of stool pathogens (Salmonella, Shigella, Vibro, Campylobacter, Yersinia, (C. difficile toxin A/B some assays). Many Types. Comments Good general purpose media. Dual swabs most convenient Good general purpose media. Yield for facultative anaerobes may be higher than from Stuarts. Best media for GC All stool specimens that can not be setup within 1 hour should be placed in Cary-Blair media CaryBlair media especially useful for Campylobacter. Recommend media with oxygen indicator. General transport media are not good for strict anaerobes. Do not refrigerate. Media that do not contain mercury or formalin are more environmentally friendly.

Amies with Charcoal Cary-Blair

Anaerobic Transport Media

Ova and Parasite media (PVA, SAF, 10% formalin, Alcohol based Ecofix) Viral Transport Media

Protozoa quickly lost in unpreserved stool

Many types

Most contain antibiotics which renders then unusable for bacterial culture.

Skin and Soft Tissue (Wound) Cultures

Collect with steel (needle aspirate or scalpel) Discourage (kurangi) the use of swabs If infection NOT suspected, DONT culture Get infected tissue or body fluid [ discourage swabs! ] -use something sharp ( syringe, scalpel, etc ) -close doesnt count *Dont culture the surface / get deep infected sample* Remove needles / send capped (tertutup) syringe with aspirate Share specimen: Microbiology-Surgical Path-Cytology ** Label specimen and site accurately ** Give appropriate history
(Matkoski C. Sharp SE, Kiska DL. Evaluation of the Q Score and Q234 Systems for cost-effective and
clinically relevant interpretation of wound cultures. J Clin Microbiol 2006;44:1869-1872)

Principle #1: The specimen must be collected with a minimum of contamination as close to site of infection as possible
Specimen Urine Culture Source of Contamination All non surgical samples become contaminated with urogenital flora during collection. Contaminating bacteria will replicate if specimen is not quickly transferred to a preservative tube or stored (4C). Improper cleaning of skin or catheter prior to drawing specimen. Transfer from SPS tube to blood culture vial. Collection from catheter. Storage and Transport Transfer urine to a Urine Preservative tube within 10 minutes of collection (good for 48 hrs. at ambient temp. Less optimal: store/transport urines at 4 C for up to 24 hrs. Ambient. Must be incubated in automated system within 12 hours. Solution/Monitor Patients must be instructed to properly cleanse the peri-urethral genital skin area prior to collection of the midstream portion of the urine stream in order to get an accurate urine culture result. Use of urine preservative tubes. Ongoing education program. Monitoring contamination rates. Limit use SPS tubes. Do not draw from catheter unless specifically requested (protocol; discard 5X cath. volume); then one culture set from catheter and one from peripheral. Education Prompt feedback to individuals or sites who collected urine for culture. Urine preservative tubes should be used when appropriate. Timely feedback to individuals who collected specimen.

Blood Culture, bacterial, mycobacterial, fungal

Principle #1: The specimen must be collected with a minimum of contamination as close to site of infection as possible
Specimen Urine Culture Source of Contamination All non surgical samples become contaminated with urogenital flora during collection. Contaminating bacteria will replicate if specimen is not quickly transferred to a preservative tube or stored (4C). Improper cleaning of skin or catheter prior to drawing specimen. Transfer from SPS tube to blood culture vial. Collection from catheter. Storage and Transport Transfer urine to a Urine Preservative tube within 10 minutes of collection (good for 48 hrs. at ambient temp. Less optimal: store/transport urines at 4 C for up to 24 hrs. Ambient. Must be incubated in automated system within 12 hours. Solution/Monitor Patients must be instructed to properly cleanse the peri-urethral genital skin area prior to collection of the midstream portion of the urine stream in order to get an accurate urine culture result. Use of urine preservative tubes. Ongoing education program. Monitoring contamination rates. Limit use SPS tubes. Do not draw from catheter unless specifically requested (protocol; discard 5X cath. volume); then one culture set from catheter and one from peripheral. Education Prompt feedback to individuals or sites who collected urine for culture. Urine preservative tubes should be used when appropriate. Timely feedback to individuals who collected specimen.

Blood Culture, bacterial, mycobacterial, fungal

Blood Cultures
Volume of blood drawn is the single most important factor influencing sensitivity. A single set for an adult blood culture consists of one aerobic and one anaerobic bottle. Optimally 10 mL of blood should be inoculated into each bottle. Volume of blood for a pediatric culture can be related to the infants weight Solitary blood cultures should be less than 5% (Arch
Pathol Lab Med. 2001 125:1290-1294)

If only enough blood can be drawn for one bottle, inoculate the aerobic bottle.

Pediatric Blood Cultures - Volume

Recommended Pediatric Blood Culture Volumes By Patient Weight Weight Weight Minimum One Two Adult (KG) of (LB) of Volume Pediatric Bottles Patient Patient (mL) Bottle (aerobic and anaerobic) <1.0 Kg 2.2 Lb. 1.0 mL Yes No 1.5-3.9 Kg 2.3-8.6 Lb. 1.5 mL Yes No 4.0-13.9 Kg 8.7-30.6 Lb 3.0 mL Yes No 14.0-24.9 30.7-54.9 10.0 mL No Yes (5 mL in Kg Lb each) >25.0 Kg >55 Lb. 20.0 Ml No Yes (10 mL in each)

URINE SPECIMEN
IN SYMPTOMATIC PATIENTS ONE SPECIMEN USSUALLY SUFFICIENT, IF ASYMPTOMATIC THREE SPECIMEN MAY BE REQUIRED THE FIRST MORNING SPECIMEN IS PREFERRED.THE FIRST PORTION OF THE URINE SHOULD BE VOIDED AS IT CONTAINS CONTAMINANTS FROM THE URETHRA. THE MID STREAM PORTION OF THE URINE SHOULD CONTAIN ANY ORGANISMS FROM THE BLADDER (kandung kemih) THE EXTERNAL AREAS OF THE GENETALIA SHOULD BE CLEANED WITH ANTIBACTERIAL SOAP PRIOR TO SPECIMEN COLLECTION IF THE SPECIMEN CANNOT BE TRANSPORTED TO THE LABORATORY IMMEDIATELY, REFRIGERATE AT 4O C SPECIMEN OLDER THAN 24 HOURS WILL NOT BE ACCEPTED

WOUND SPECIMENS

REMOVE AS OF MUCH THE SKIN FLORA WITH THE USE OF A SKIN DESINFECTANT THE SPECIMEN SHOULD COLECTED FROM THE ADVANCING MARGIN OF THE LESION OR THE ABSCESS WALL IF THE SPECIMEN CANNOT BE TRANSPORTED TO THE LABORATORY IMMEDIATELY, REFRIGERATE AT -4O C

GENETAL SPECIMEN

MUCOUS AND/OR SECRETIONS SHOULD BE REMOVED FROM SURFACE WITH A STERILE SOAP A SECOND SWAB IS USED TO COLECT SAMPLE FROM SELECTED SIDE

SPUTUM SPECIMEN

WHEN REQUIRED, THE FIRST EARLY MORNING IS PREFERRED THE MOUTH SHOULD BE RINSED OUT (kumur) WITH WATER SPECIMEN SHOULD BE COLLECTED DIRECTLY INTO STERILE CONTAINER

THROAT SPECIMEN
UPPER RESPIRATORY INFECTIONS ARE CAUSED BY A MIXTURE OF BACTERIAL DAN VIRAL PATHOGEN ACUT PHARYNGITIS AND TONSILLITIS ARE MOST OFTEN CAUSED BY BACTERIAL PATHOGENS USING THE TONGUE BLADE PRESS DOWN ON THE TONGUE AND SWAB THE TONSIL AND POSTERIOR PHARYNX DO NOT TOUCH THE SWAB ON ANY OTHE ORAL SURFACE

NASAL SPECIMEN
THE SPECIMEN OF CHOISE IS FROM A REGION AT LEAST 1 CM INSIDE THE NARES THE SWAB SHOULD BE INSERTED AT LEAST 1 CM INTO THE NARES THE SWAB SHOULD ROTATED AGAINST THE MEMBRANE AT LEAST 10 TO 15 SECOND

FUNGAL SPECIMEN
1. SKIN SCRAPING: CLEAN THE SURFACE OF SKIN WITH ALKOHOL 70% , SCRAPE MATERIAL FROM ACTIVE MARGIN OF THE LESION COLLECTING IN TO CLEAN CONTAINER 2. HAIR: REMOVE AT LEAST 10 TO 12 AFFECTED HAIR WITH FORCEPS. IF AVAILABLE SELECT HAIRS WITH THE AID OF A WOODS LAMP. NOTE THAT NOT ALL DERMATOPHYTE WILL FLUOREC 3. NAIL (kuku) : REMOVE ANY NAIL POLISH (OBAT GOSOK) FROM NAIL. COLLECT DEBRIS UNDER THE AFFECTED PORTION OF THE NAIL. IF POSIBLE COLLECT SCRAPING FROM DEEPER DISEASE PORTION OF THE NAIL

Cultures That Should Include a Gram Stain

CSF or sterile body fluid (cytospin) Eye Purulent discharge (KOTORAN) Sputum or transtracheal aspirate All surgical specimens Tissue Urethral exudates (male only, intracellular gonococcus)) Vaginal specimens Wounds

Criteria For Rejection of Microbiological Specimens

Criteria for rejection must be readily available and laboratory specific Unlabeled or improperly labeled specimen Prolonged storage or transport Improper or damaged container Specimen received in fixative Oropharyngeal contaminated sputum Duplicate specimens (stools, sputum) within a 24 hour period. Exceptions cleared by the laboratory Specimens unsuitable for culture request (anaerobic culture from not acceptable source, urine from Foley catheter) Dry Swab 24-hr collection of urine or sputum for AFB or fungal culture Other criteria specific to your laboratory