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利用单克隆抗体快速检测和鉴定乙型流感病毒的两个谱系毒株

Rapid detection and identification of two lineages of influenza B strains with monoclonal antibodies.

作者信息

Nakagawa N, Maeda A, Kase T, Kubota R, Okuno Y

机构信息

Department of Public Health, Osaka Prefectural Institute of Public Health, Japan.

出版信息

J Virol Methods. 1999 Apr;79(1):113-20. doi: 10.1016/s0166-0934(99)00015-4.

DOI:10.1016/s0166-0934(99)00015-4
PMID:10328540
Abstract

Monoclonal antibodies (Mabs) against influenza B virus were obtained by immunizing mice with B/Nagasaki/1/87, one of the strains of the B/Victoria group. Immunoprecipitation analysis revealed that individual Mabs precipitated the nucleoprotein (NP), the matrix protein (M) or the hemagglutinin protein (HA). By using these Mabs by the peroxidase-antiperoxidase (PAP) staining method, a rapid detection and identification method for influenza B virus was established. Monolayers of Madin-Darby canine kidney cells in microplates were infected with each-strain and incubated for about 24 h, and then were subjected to the PAP staining method using the Mabs as the first antibody. Influenza B virus strains are classified into two major phylogenetic trees, the B/Victoria group and the B/Yamagata group. When anti-NP and anti-M antibodies were used in the PAP staining method, all 13 influenza B virus strains isolated from clinical specimens between 1940 and 1994 were detected regardless of the antigenic drift of the influenza virus. On the other hand, several anti-HA Mabs which reacted specifically with the strains of the B/Victoria group, did not react with any strain of the B/Yamagata group. In the 1996/97 influenza season in Osaka Prefecture in Japan, two antigenically distinct groups of influenza B virus strains were isolated. They belonged to different phylogenetic trees and were clearly distinguishable by the PAP staining method with anti-HA Mabs.

摘要

用B/维多利亚系的一个毒株B/长崎/1/87免疫小鼠,获得了抗乙型流感病毒的单克隆抗体。免疫沉淀分析表明,单个单克隆抗体沉淀核蛋白(NP)、基质蛋白(M)或血凝素蛋白(HA)。通过使用这些单克隆抗体采用过氧化物酶抗过氧化物酶(PAP)染色法,建立了一种快速检测和鉴定乙型流感病毒的方法。在微孔板中用各毒株感染麦迪逊-达比犬肾细胞单层,并孵育约24小时,然后以单克隆抗体作为一抗采用PAP染色法。乙型流感病毒毒株分为两个主要的系统发育树,即B/维多利亚系和B/山形系。当在PAP染色法中使用抗NP和抗M抗体时,1940年至1994年间从临床标本中分离出的所有13株乙型流感病毒毒株均被检测到,而不考虑流感病毒的抗原漂移。另一方面,几种与B/维多利亚系毒株特异性反应的抗HA单克隆抗体,与B/山形系的任何毒株均无反应。在日本大阪府1996/97流感季节,分离出两组抗原性不同的乙型流感病毒毒株。它们属于不同的系统发育树,通过用抗HA单克隆抗体的PAP染色法可明显区分。

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