1. Processing of Clinical Specimen for Immunofluorescence staining & Viral culture Exercise II – Day 4 Virology Section Department of Microbiology JIPMER XIII NATIONAL WORKSHOP ON “DIAGNOSTIC METHODS IN CLINICAL MICROBIOLOGY” 8 th – 11 th October, 2014

2. Introduction Viral detection methods: Staining  Direct staining  Immunofluorescence staining Culture Cell culture Animal / embryonated egg inoculation Enzyme immunoassays PCR methods XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER 2

3. Objective To observe and understand the steps in processing a clinical specimen for immunofluorescence staining and viral culture XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER 3

4. Steps – Processing of specimen Sample receipt V ortex & aliquot Centrifug e XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER 4 Use for Viral culture Use for Immunofluorescence staining

5. IFA XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER 5

6. Positive IFA- Adenovirus Negative IFA XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER 6

7. Detection of virus growth in cell cultures  Cytopathic effect  Metabolic inhibition  Hemadsorption  Interference  Transformation  Immunofluorescence XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER 7

8. Normal cell line – Hep2 CPE- Adenovirus XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER Grape like clusters of round cells Polygonal cells

9. Thank you XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER