Guizhou Provincial People's Hospital, Medical College, Guizhou University, No. 83 Zhongshan Dong Road, Guiyang, 550002, Guizhou Province, China.
Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12- 4, Sakamoto, Nagasaki, 852-8523, Japan.
Virol J. 2018 Nov 22;15(1):178. doi: 10.1186/s12985-018-1071-y.
Rift Valley Fever (RVF) is a mosquito-borne viral zoonosis. To detect RVF virus (RVFV) infection, indirect immunoglobulin G (IgG) and immunoglobulin M (IgM) enzyme linked immunosorbent assays (ELISAs) which utilize recombinant RVFV nucleocapsid (RVFV-N) protein as assay antigen, have reportedly been used, however, there is still a need to develop more sensitive and specific methods of detection.
RVFV-N protein was expressed in Escherichia coli (E. coli) and purified by histidine-tag based affinity chromatography. This recombinant RVFV-N (rRVFV-N) protein was then used as antigen to develop an IgG sandwich ELISA and IgM capture ELISAs for human sera. Ninety six serum samples collected from healthy volunteers during the RVF surveillance programme in Kenya in 2013, and 93 serum samples collected from RVF-suspected patients during the 2006-2007 RVF outbreak in Kenya were used respectively, to evaluate the newly established rRVFV-N protein-based IgG sandwich ELISA and IgM capture ELISA systems in comparison with the inactivated virus-based ELISA systems.
rRVFV-N protein-based-IgG sandwich ELISA and IgM capture ELISA for human sera were established. Both the new ELISA systems were in 100% concordance with the inactivated virus-based ELISA systems, with a sensitivity and specificity of 100%.
Recombinant RVFV-N is a safe and affordable antigen for RVF diagnosis. Our rRVFV-N-based ELISA systems are safe and reliable tools for diagnosis of RVFV infection in humans and especially useful in large-scale epidemiological investigation and for application in developing countries.
裂谷热（RVF）是一种由蚊子传播的病毒性人畜共患病。为了检测裂谷热病毒（RVFV）感染，已经报道了使用间接免疫球蛋白 G（IgG）和免疫球蛋白 M（IgM）酶联免疫吸附试验（ELISA），这些试验使用重组 RVFV 核衣壳（RVFV-N）蛋白作为检测抗原，然而，仍然需要开发更敏感和特异的检测方法。
RVFV-N 蛋白在大肠杆菌（E. coli）中表达，并通过组氨酸标签基于亲和层析进行纯化。然后，将这种重组 RVFV-N（rRVFV-N）蛋白用作抗原，开发用于人血清的 IgG 夹心 ELISA 和 IgM 捕获 ELISA。分别使用 2013 年肯尼亚裂谷热监测计划中从健康志愿者收集的 96 份血清样本和 2006-2007 年肯尼亚裂谷热爆发期间从裂谷热疑似患者收集的 93 份血清样本，来评估新建立的基于 rRVFV-N 蛋白的 IgG 夹心 ELISA 和 IgM 捕获 ELISA 系统与基于灭活病毒的 ELISA 系统的比较。
建立了用于人血清的 rRVFV-N 蛋白基于 IgG 的夹心 ELISA 和 IgM 捕获 ELISA。这两种新的 ELISA 系统与基于灭活病毒的 ELISA 系统完全一致，灵敏度和特异性均为 100%。
重组 RVFV-N 是 RVF 诊断的一种安全且经济实惠的抗原。我们基于 rRVFV-N 的 ELISA 系统是用于诊断人类 RVFV 感染的安全可靠工具，尤其在大规模流行病学调查和发展中国家的应用中非常有用。
