Ziegler T, Hall H, Sánchez-Fauquier A, Gamble W C, Cox N J
Influenza Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 30333.
J Clin Microbiol. 1995 Feb;33(2):318-21. doi: 10.1128/jcm.33.2.318-321.1995.
A rapid culture assay which allows for the simultaneous typing and subtyping of currently circulating influenza A(H1N1), A(H3N2), and B viruses in clinical specimens was developed. Pools of monoclonal antibodies (MAbs) against influenza A and B viruses and MAbs HA1-71 and HA2-76, obtained by immunizing mice with the denatured hemagglutinin subfragments HA1 and HA2 of influenza virus A/Victoria/3/75, were used for immunoperoxidase staining of antigens in infected MDCK cells. MAb HA1-71 reacted exclusively with influenza A viruses of the H3 subtype, while MAb HA2-76 reacted with subtypes H1, H3, H4, H6, H8, H9, H10, H11, and H12, as determined with 78 human, 4 swine, and 10 avian influenza virus reference strains subtyped by the hemagglutination inhibition test. To determine if the technique can be used as a rapid diagnostic test, 263 known influenza virus-positive frozen nasal or throat swabs were inoculated into MDCK cells. After an overnight 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 31 and 113 specimens, respectively. The subtypes of 10 influenza A virus-positive specimens could not be determined because they contained too little virus. Influenza B viruses were detected in 84 specimens, and 25 specimens were negative. We conclude that this assay is a rapid, convenient, non-labor-intensive, and relatively inexpensive test for detecting, typing, and subtyping influenza viruses in clinical specimens.
开发了一种快速培养检测方法，可对临床标本中当前流行的甲型（H1N1）、甲型（H3N2）和乙型流感病毒同时进行分型和亚型鉴定。使用针对甲型和乙型流感病毒的单克隆抗体（MAb）池以及通过用甲型流感病毒A/维多利亚/3/75的变性血凝素亚片段HA1和HA2免疫小鼠获得的单克隆抗体HA1-71和HA2-76，对感染的MDCK细胞中的抗原进行免疫过氧化物酶染色。单克隆抗体HA1-71仅与H3亚型的甲型流感病毒反应，而单克隆抗体HA2-76与H1、H3、H4、H6、H8、H9、H10、H11和H12亚型反应，这是通过血凝抑制试验对78株人、4株猪和10株禽流感病毒参考毒株进行亚型鉴定确定的。为了确定该技术是否可用作快速诊断测试，将263份已知流感病毒阳性的冷冻鼻拭子或咽拭子接种到MDCK细胞中。过夜培养后，固定细胞并通过免疫过氧化物酶染色检测病毒抗原。分别在31份和113份标本中检测到H1和H3亚型的甲型流感病毒。10份甲型流感病毒阳性标本的亚型无法确定，因为它们含有的病毒太少。在84份标本中检测到乙型流感病毒，25份标本为阴性。我们得出结论，该检测方法是一种快速、便捷、无需大量劳动力且相对廉价的检测临床标本中流感病毒、进行分型和亚型鉴定的方法。
