Huang Yi, Zhu Youjie, Yang Mengshi, Zhang Zhenqing, Song Donglin, Yuan Zhiming
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Virol Sin. 2014 Dec;29(6):372-80. doi: 10.1007/s12250-014-3512-0. Epub 2014 Dec 15.
Full-length nucleoproteins from Ebola and Marburg viruses were expressed as His-tagged recombinant proteins in Escherichia coli and nucleoprotein-based enzyme-linked immunosorbent assays (ELISAs) were established for the detection of antibodies specific to Ebola and Marburg viruses. The ELISAs were evaluated by testing antisera collected from rabbit immunized with Ebola and Marburg virus nucleoproteins. Although little cross-reactivity of antibodies was observed in anti-Ebola virus nucleoprotein rabbit antisera, the highest reactions to immunoglobulin G (IgG) were uniformly detected against the nucleoprotein antigens of homologous viruses. We further evaluated the ELISA's ability to detect antibodies to Ebola and Marburg viruses using human sera samples collected from individuals passing through the Guangdong port of entry. With a threshold set at the mean plus three standard deviations of average optical densities of sera tested, the ELISA systems using these two recombinant nucleoproteins have good sensitivity and specificity. These results demonstrate the usefulness of ELISA for diagnostics as well as ecological and serosurvey studies of Ebola and Marburg virus infection.
埃博拉病毒和马尔堡病毒的全长核蛋白在大肠杆菌中表达为带His标签的重组蛋白,并建立了基于核蛋白的酶联免疫吸附测定(ELISA)法,用于检测埃博拉病毒和马尔堡病毒特异性抗体。通过检测用埃博拉病毒和马尔堡病毒核蛋白免疫的兔子所采集的抗血清,对ELISA法进行了评估。虽然在抗埃博拉病毒核蛋白兔抗血清中观察到抗体的交叉反应很少,但针对同源病毒的核蛋白抗原,均检测到对免疫球蛋白G(IgG)的最高反应。我们进一步使用从通过广东入境口岸的人员采集的人血清样本,评估了ELISA法检测埃博拉病毒和马尔堡病毒抗体的能力。将阈值设定为所检测血清平均光密度的平均值加三个标准差,使用这两种重组核蛋白的ELISA系统具有良好的敏感性和特异性。这些结果证明了ELISA法在埃博拉病毒和马尔堡病毒感染的诊断以及生态学和血清学调查研究中的有用性。