Dr Mohit Bhatia Assistant Professor Department of Microbiology AIIMS Rishikesh SPECIMENS Upper respiratory tract specimens 1 Deep nostrils Nasal swab 2 Nasopharynx Nasopharyngeal swab ID: 997853
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1. LABORATORY DIAGNOSIS OF SWINE FLUDr. Mohit BhatiaAssistant ProfessorDepartment of MicrobiologyAIIMS, Rishikesh
2. SPECIMENSUpper respiratory tract specimens: (1) Deep nostrils (Nasal swab)(2) Nasopharynx (Nasopharyngeal swab)(3) Nasopharyngeal aspirate(4) Bronchial aspirate(5) Throat swabAcute and convalescent serum specimens should be used for the detection of rising antibody titres
3. POSTMORTEM SAMPLINGLung: all lobes, including affected areas Proximal and distal trachea Liver Brain Kidneys Nasopharynx (this is best done from a supratentorial approach with removal en bloc).
4. DIAGNOSTIC TESTS
5. VIRAL CULTUREEmbryonated eggsMammalian cell lines:(1) Madin Darby canine kidney (MDCK)(2) A549(3) Mink lung epithelial cell line (Mv1Lu)(4) Rhesus monkey kidney (LLC MK2) (5) Buffalo green monkey kidney (BGMK)(6) Primary cell lines: Rhesus monkey kidney (RhMK) or African green monkey kidney (AGMK)
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7. SHELL VIAL CULTURE
8. DIRECT FLUORESCENT ANTIBODY TEST (DFA)Mainly intended for the detection and differentiation of influenza A and B viruses. Not useful for subtyping influenza A viruses.
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10. SEROLOGICAL ASSAYSHemagglutination inhibition assay (HAI)Microneutralization or virus neutralization assay (VN)Single radial hemolysis (SRH)Complement fixation assayEnzyme linked immunoabsorbant assay (ELISA)Western blotting.
11. HEMAGGLUTINATION INHIBITION ASSAY
12. VIRUS NEUTRALIZATION ASSAY
13. SINGLE RADIAL HEMOLYSISSRH is a technique commonly used to determine the induction of antibodies following natural infection or vaccination. It is used to measure complement-mediated hemolysis induced by antigen-antibody complexes.SRH is considered more sensitive than the HAI assay and does not require pretreatment of the serum to inactivate non-specific inhibitors.
14. COMPLEMENT FIXATION TEST
15. ELISA
16. RAPID INFLUENZA DIAGNOSTIC TESTS (RIDTs)
17. LAB ON A CHIP/MICROCHIP DEVICES
18. NUCLEIC ACID BASED TESTS (NATs)Reverse Transcriptase Polymerase Chain Reaction (RT-PCR)Loop-Mediated Isothermal Amplification-Based Assay (LAMP)Simple Amplification-Based Assay (SAMBA)Nucleic Acid Sequence-Based Amplification (NASBA)Microarray-Based Approaches
19. REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION (RT-PCR)
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21. LOOP-MEDIATED ISOTHERMAL AMPLIFICATION-BASED ASSAY (LAMP)
22. SIMPLE AMPLIFICATION-BASED ASSAY (SAMBA)
23. NUCLEIC ACID SEQUENCE-BASED AMPLIFICATION (NASBA)
24. MICROARRAY-BASED APPROACHES
25. NUCLEIC ACID SEQUENCING SANGER SEQUENCINGNEXT GENERATION SEQUENCING (NGS)
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