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

临床标本中流感病毒的快速检测、分型和亚型分析

基本信息

批准号：
06670443
负责人：
MAEDA Akiko
金额：
$ 1.15万
依托单位：
OSAKA PREFECTURAL INSTITUTE OF PUBLIC HEALTH
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

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.

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) 和乙型流感病毒进行同步分型和亚型分析，针对甲型 (H1N1) 流感：C179 和甲型 (H3N2) 流感的单克隆抗体： F49和抗乙型流感多克隆抗体用于抗原感染的MDCK细胞的免疫过氧化物酶染色。 Eecept 2 株、单克隆抗体和多克隆抗体仅与甲型流感病毒和乙型流感病毒发生反应，这是通过血凝抑制试验对 160 种流感病毒参考株进行分型或亚型确定的。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型和AH3型流感混合感染。为了确定该技术是否可以用作快速诊断测试，在1993/94和1994/95流感流行中，将72和80个临床标本接种到MDCK细胞中。孵育40小时后，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27（1994/95）和25（1993/94）样本中检测到H1和H3亚型甲型流感病毒。 8份AH3型流感病毒和7份AH1型流感病毒阳性标本因病毒含量太少而无法确定亚型。我们的结论是，该测定是一种快速且相对便宜的检测方法，用于检测临床标本中的流感病毒、对其进行分型和分型。