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West Nile Virus 2002 Laboratory Perspectives Laboratories Branch MOHLTC. Pauline George. Overview. Test procedures WNV Statistics Laboratory Challenges. Principle of HI Test. Viral Antigen + Goose RBCs = Agglutination
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West Nile Virus 2002Laboratory PerspectivesLaboratories Branch MOHLTC Pauline George
Overview Test procedures WNV Statistics Laboratory Challenges
Principle of HI Test Viral Antigen + Goose RBCs = Agglutination Serum (Abs) + VAg + GRBCs = No Agglut/Reactive Serum ( no Abs) + VAg + GRBCs = Agglut/Nonreactive
Test Procedures-Single Serum HI Test Process • Day 1 Extraction • Day 2 Absorption • Day 2 Test Screen/Titrate • Day 3 Add RBCs • Day 3 Read
Confirmatory TestWinnipeg Federal Lab • 2 week TAT • Plaque Reduction Neutralization test Testing on paired serum against likely viruses Presence/absence neutralizing antibody • PCR Tissue/CSF
WVN CasesSummary (to Nov 4, 2002) • Confirmed * (PRNT) 49 • Did not confirm *(PRNT) 9 • Probable * (HI) 89 • Suspect ^ 176 * seroconversion ^ seropositive
Laboratory Challenges 2002 • Substantially increased test volumes • Staffing resources • Reagent Availability • Equipment • Antigenic Variation • Increased incoming/outgoing telephone calls • Other routine tests • Training • TDG • Space Allocation
Future Challenges 2002 and Beyond • Funding • Staffing • In-house tests vs Commercial Tests Availability? • Case criteria • Data Analysis • Training • Confirmatory Test Availability • Antigen consistency • Antigen source/cost?
Future Challenges2002 and Beyond cont’d • For confirmatory testing in Ontario • Requires Level 3 Biosafety lab • PRNT confirmatory assay • PCR • Culture
Acknowledgements OPHL Arboviruses & Rickettsiae Laboratory Staff Dr. M. Fearon, I Guglielmi, E Cheung, Canadian Science Centre for Human & Animal Health , Winnipeg Viral Zoonotic Staff Dr. M. Drebot