Jiangxi Province Key Laboratory of Preventive Medicine, School of Public Health, Nanchang University, Nanchang, P. R. China.
National Institute for Viral Disease Control and Prevention, China CDC, Key Laboratory for Medical Virology, National Health Commission, Beijing, P. R. China.
J Med Virol. 2019 Jun;91(6):1168-1171. doi: 10.1002/jmv.25408. Epub 2019 Feb 4.
Avian H7N9 subtype influenza virus infects human with high case-fatality rate since it emerged in 2013. Although the vaccination has been rapidly used in poultry due to the emergence of highly pathogenic strain, this virus remains prevalent in this region. Thus, rapid diagnosis both in poultry and human clinic is critically important for the control and prevention of H7N9 infection. In this study, a batch of H7 subtype-specific monoclonal antibodies (mAbs) were developed and a pair of mAb, 2B6, and 5E9 were used to establish a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) to quantify H7 protein and detect influenza A virus baring H7 subtype HA. The lowest detection limit for the recombinant H7 protein was 10 ng/mL and 0.5 HAU/50 μL of A/Guangdong/17SF003/2016(H7N9), 2 HAU/50 μL of A/Netherlands/219/2003(H7N7) and A/Anhui/1/2013(H7N9) for live virus, respectively. The ELISA could not only detect the prevailing H7N9 virus, but also antigenic drift H7 subtype viruses, showing excellent sensitivity and high specificity. Hence, it could serve as a valuable approach to diagnose H7 subtype virus which showed great potential to cause pandemic, as well as antigen quantification.
自 2013 年出现以来，禽源 H7N9 亚型流感病毒感染人类的病死率很高。虽然由于高致病性毒株的出现，疫苗已在禽类中迅速使用，但该病毒在该地区仍然流行。因此，快速诊断禽类和人类临床的 H7N9 感染对于控制和预防 H7N9 感染至关重要。本研究中开发了一批 H7 亚型特异性单克隆抗体（mAbs），并使用一对 mAb（2B6 和 5E9）建立了双抗体夹心酶联免疫吸附试验（ELISA）来定量 H7 蛋白并检测携带 H7 亚型 HA 的甲型流感病毒。重组 H7 蛋白的最低检测限为 10ng/mL，活病毒的 A/Guangdong/17SF003/2016（H7N9）为 0.5 HAU/50 μL，A/Netherlands/219/2003（H7N7）和 A/Anhui/1/2013（H7N9）为 2 HAU/50 μL。该 ELISA 不仅可以检测流行的 H7N9 病毒，还可以检测抗原漂移的 H7 亚型病毒，显示出优异的灵敏度和高度的特异性。因此，它可以作为一种有价值的方法来诊断可能引起大流行的 H7 亚型病毒，以及进行抗原定量。
