H5N1-SeroDetect EIA and Rapid Test: a Novel Differential Diagnostic Assay for Serodiagnosis of H5N1 Infections and Surveillance
H5N1-SeroDetect EIA and Rapid Test: a Novel Differential Diagnostic Assay for Serodiagnosis of H5N1 Infections and Surveillance
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DOI：
10.1128/jvi.06023-11
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发表时间：
2011-12-01
影响因子：
5.4
通讯作者：
Golding, Hana
中科院分区：
文献类型：
--
作者：
Khurana, Surender;Sasono, Pretty;Golding, Hana
高致病性（HP）H5 N1流感病毒在野鸟中的持续进化并传播给家禽和人类构成了大流行的威胁。迫切需要一种简单快速的血清学诊断测定法，它可以区分季节性和H5 N1毒株的抗体，并可以提供不依赖于病毒分离和核酸技术的监测工具。在这里，我们描述了H5 N1 SeroDetect酶联免疫吸附试验（ELISA）的建立以及基于HA 2（488-516）、PB 1-F2（2-75）和M2 e（2-24）中三种肽的快速检测试验，这些肽在H5 N1毒株中高度保守。这些肽是通过使用全基因组片段噬菌体展示文库（GFPDLs）对越南H5 N1流感幸存者进行抗体库分析来鉴定的。迄今为止，这两个平台在检测越南患者感染H5 N1（进化枝1和进化枝2.3.4）方面表现出高水平的灵敏度和特异性，最早可在感染后7天至数年内检测到。在越南和美国，未感染H5 N1病毒的个体，包括接种季节性流感疫苗或确诊季节性病毒感染的受试者，在H5 N1-SeroDetect检测试剂盒中没有反应。此外，来自接种H5 N1亚单位疫苗的具有中等抗H5 N1中和抗体滴度的个体的血清在H5 N1-SeroDetect ELISA或快速测试测定中没有阳性反应。简单的H5 N1-SeroDetect ELISA和快速检测可以为大规模监测人类和鸟类可能接触H5 N1毒株提供重要工具。
Continuing evolution of highly pathogenic (HP) H5N1 influenza viruses in wild birds with transmission to domestic poultry and humans poses a pandemic threat. There is an urgent need for a simple and rapid serological diagnostic assay which can differentiate between antibodies to seasonal and H5N1 strains and that could provide surveillance tools not dependent on virus isolation and nucleic acid technologies. Here we describe the establishment of H5N1 SeroDetect enzyme-linked immunosorbent assay (ELISA) and rapid test assays based on three peptides in HA2 (488-516), PB1-F2 (2-75), and M2e (2-24) that are highly conserved within H5N1 strains. These peptides were identified by antibody repertoire analyses of H5N1 influenza survivors in Vietnam using whole-genome-fragment phage display libraries (GFPDLs). To date, both platforms have demonstrated high levels of sensitivity and specificity in detecting H5N1 infections (clade 1 and clade 2.3.4) in Vietnamese patients as early as 7 days and up to several years postinfection. H5N1 virus-uninfected individuals in Vietnam and the United States, including subjects vaccinated with seasonal influenza vaccines or with confirmed seasonal virus infections, did not react in the H5N1-SeroDetect assays. Moreover, sera from individuals vaccinated with H5N1 subunit vaccine with moderate anti-H5N1 neutralizing antibody titers did not react positively in the H5N1-SeroDetect ELISA or rapid test assays. The simple H5N1-SeroDetect ELISA and rapid tests could provide an important tool for large-scale surveillance for potential exposure to HP H5N1 strains in both humans and birds.