RAPID TEST FOR DETECTION,TYPING AND SUBTYPING INFLUENZA VIRUSES IN CLINICAL SPECIMENS

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A rapid culture assay whitch allows for the simultaneous typing and subtyping of currently circulating influenza A (H1N1), A (H3N2) and B viruses in clinical specimens was developed, monoclonal antibodies against influenza A (H1N1) : C179 and A(H3N2) : F49 and polyclonal antibody against influenza B,were used for immunoperoxidase staining of antigens infected MDCK cells. Eecept 2 strains, monoclonal antibodies and polyclonal antibody react exclusively with influenza A viruses and influenza B Virus, as determined with 160 influenza virus reference strains typed or subtyped by the hemagglutination-inhibition test. 2 strains were proved the mixed infection of influenza AH1 and AH3 by immunoperoxidase staining of antigens infected MDCK cells. To determine if the technique can be used as a rapid diagnostic test, on 1993/94 and 1994/95 epidemics of influenza, 72 and 80 clinical specimens were inoculated into MDCK cells. After 40 hours incubation, the cells were fixed and viral antigens were detected by immunoperoxidase staining. Influenza A viruses of the H1 and H3 subtypes were detected in 27 (1994/95) and 25(1993/94) specimems, respectively. The subtypes of 8 influenza AH3 and 7 influenza AH1 virus-positive specimems could not be determind because they conteined too little virus. We conclude that this assay is a rapid and relatively inexpensive test for detection, typing and subtyping influenza viruses in clinical specimens.