Direct demonstration of viral Ag or nucleic acid EM Serological demonstration of Florescent Ab. Ab to the virus. Double I.D. I.F. Probing ELISA Isolation and identification PCR NT of the virus. HAI T.C. Chick embryo SERUM Lab. animal CLINICAL SPECIMEN
EM.picture of viruses
2) Fluorescent-antibody test
Viral antigen in smears detected by direct immunofluorescent technique (DIF). Specific fluorescencet stain with standard antiviral serum
Examined by UV microscope.
Immunofluorescent technique
3) Double immunodiffusion
V. antigen can be detected by precipitation against antiviral serum in agar gel Rapid method and takes 2-6 hours
4-ELISA
Double antibody sandwitch technique for detecting antigen.
5-Nucleic acid probe: DNA probes are pieces of nucleic acid labeled in some fashion that can find to structures of DNA that have complementary sequences. So unknown organism can be identified.
Cytological examination:
Demonstration of characteristic cytological changes from smears of infected tissues. Inclusion bodies ( IB ),multinucleated( syncetial ) giant cells are the main changes. IB are masses of viral particles seen in nucleus or cytoplasm; intranuclear or intracytoplasmic,either acidophilic or basophilic in nature. Diseases as HSV, VZ virus and rabies produce cytologiccal changes .
B-Virus isolation
Specimen: Body secretions or excretions, or from the site of lesion. Taken on a swab to preserve the virus on transport media (TC. Media) Specimen reach the lab as fast as possible. Specimen must be kept cold. Antibiotics and antifungal are added to the specimen in the lab, avoid bact. contamination. Specimen is centrifuged to deposit the bact.
Tissue culture
Chick embryo
Laboratory animals
Virus TC-cell line incubated signs of growth identification and typing of virus.
1-Tissue culture: TC- Cells from man or animal are grown as a single layer (mono-layer) on the wall of the tubes or on one side of flat bottle. Cells are incubated at 37C. Suspended in tissue culture media
2) Haemadsorption Seen with haemagglutinating viruses which mature at the cell surface. Virus infected cell + Erythrocytes eryth. Adherent to the infected cell.
3) Haemagglutination
Haemagglutination test is done on the cell fluid to detect the presence of haemagglutinating virus.
4)Interference
Haemagglutination pattern.
5) Immunofluorescence
Vs antigen is detected in infected cell by DIF.
2. Chick embryo
Fertile hen`s egg
Used before the advent of tissue culture. Susceptible to fewer no. of viruses.
C- Serological Diagnosis:
Demonstration of virus antibody in patient serum. A diagnosis of recent infection depend on: 1-Detection of IgM. 2-Rising titre: Increase in level of antibody titre; at least four fold rise. 3-High stationary titre of antibody.
The ELISA test in a microplate. Microtitre plates of special quality and with flat bottomed wells are used. The colour produced is the basis for interpretation of the reaction, which is read by a photometer. The four rows on the left are the controls.