Development of a novel rapid immunochromatographic test specific for the H5 influenza virus
Development of a novel rapid immunochromatographic test specific for the H5 influenza virus
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DOI：
10.1016/j.jviromet.2011.02.007
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发表时间：
2011-05-01
影响因子：
3.1
通讯作者：
Kida, Hiroshi
中科院分区：
文献类型：
--
作者：
Miyagawa, Eiji;Kogaki, Hiroyuki;Kida, Hiroshi
用灭活的A/duck/北海道/Vac-1/04（H5 N1）免疫BALB/C小鼠，获得3株抗H5流感病毒单克隆抗体（mAb），分别为IFH 5 -26、IFH 5 -115和IFH 5 -136。通过使用重组血凝素（HA）的蛋白质印迹法，发现这些mAb特异性识别流感H5亚型的血凝素（HA）表位;然而，这些mAb对A/duck/北海道/84/02（H5 N3）或A/Puerto Ric/8/34（H1 N1）没有中和活性。这些mAb的每个表位是构象表位，其由位于IFH 5 -115的46至60个氨基酸（aa）和312至322 aa之间、IFH 5 -136的101至113 aa和268至273 aa之间以及IFH 5 -26的61至80 aa和290至300 aa之间的区域形成。表位位于血凝素1（HA 1）受体区和α-螺旋结构之间的环区。甲型流感病毒H5特异性快速免疫层析检测试剂盒作为固相抗体/碱性磷酸酶偶联mAb以以下三种组合进行检测：IFH 5 -26/IFH 5 -115、IFH 5 -136/IFH 5 -26和IFH 5 -136/IFH 5 -115。在每个组合中，仅检测到甲型流感H5亚型。为了有效的临床应用，开发了与H5 HA特异性mAb（IFA 5 -26和IFA 5 -115）和甲型流感NP NP特异性mAb（FVA 2 -11）组合的快速双重鉴别免疫层析检测试剂盒。双重鉴别免疫层析检测试剂盒检测甲型流感病毒H5亚型为H5线阳性，同时检测所有甲型流感病毒亚型为A线阳性。双重鉴别免疫层析检测试剂盒可用于区分高致病性禽流感A H5 N1病毒和季节性流感A病毒，以及用于确认人、禽和哺乳动物宿主中的流感感染状况。(C)2011爱思唯尔有限公司版权所有。
Three anti-H5 influenza virus monoclonal antibody (mAb) clones, IFH5-26, IFH5-115 and IFH5-136, were obtained by immunising a BALB/C mouse with inactivated A/duck/Hokkaido/Vac-1/04 (H5N1). These mAbs were found to recognise specifically the haemagglutinin (HA) epitope of the influenza H5 subtypes by western blotting with recombinant HAs; however, these mAbs have no neutralising activity for A/duck/Hokkaido/84/02 (H5N3) or A/Puerto Ric/8/34 (H1N1). Each epitope of these mAbs was a conformational epitope that was formed from the regions located between 46 to 60 amino acids (aa) and 312 to 322 aa for IFH5-115, from 101 to 113 aa and 268 to 273 aa for IFH5-136 and from 61 to 80 aa and 290 to 300 aa for IFH5-26. The epitopes were located in the loop regions between the receptor region and alpha-helix structure in haemagglutinin 1 (HA1). Influenza A virus H5-specific rapid immunochromatographic test kits were tested as solid phase antibody/alkaline phosphate-conjugated mAb in the following three combinations: IFH5-26/IFH5-115, IFH5-136/IFH5-26 and IFH5-136/IFH5-115. In every combination, only influenza A H5 subtypes were detected. For effective clinical application, rapid dual discrimination immunochromatographic test kits in combination with H5 HA-specific mAb, IFA5-26 and IFA5-115 and the influenza A NP NP-specific mAb, FVA2-11, were developed. The dual discrimination immunochromatographic tests kits detected influenza A virus H5 subtypes as H5 line-positive and all influenza A subtypes as A line-positive simultaneously. The dual discrimination immunochromatographic test kits may be useful for discriminating highly pathogenic avian influenza A H5N1 viruses from seasonal influenza A virus, as well as for confirming influenza infection status in human, avian and mammalian hosts. (C) 2011 Elsevier B.V. All rights reserved.