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

建立了一种可同时对临床标本中流行的甲型H1N1流感病毒、甲型H3N1流感病毒和乙型流感病毒进行分型和分型的快速培养方法，用抗甲型H1N1流感病毒C179和抗甲型H3N1流感病毒A(H3N2)：F49的单抗和抗B型流感的多克隆抗体对感染的MDCK细胞进行免疫过氧化物酶染色。用血凝抑制试验对160株流感病毒参比毒株进行鉴定，结果表明，除2株外，单抗和多克隆抗体均能与甲型和乙型流感病毒发生特异性反应。对感染MDCK细胞的抗原进行免疫过氧化物酶染色，证实2株为AH1和AH3混合感染。为了确定该技术是否可以作为一种快速诊断试验，在1993/94年和1994/95年流感流行期间，将72份和80份临床标本接种到MDCK细胞中。孵育40h后，固定细胞，免疫过氧化物酶染色检测病毒抗原。在27份(1994/95)和25份(1993/94)样本中分别检测到H1型和H3型甲型流感病毒。8份AH3型流感和7份AH1型流感病毒阳性标本因病毒含量太少而无法确定其亚型。我们得出结论，对于临床标本中流感病毒的检测、分型和分型，该方法是一种快速且相对便宜的检测方法。

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.