Huang Jiansheng, Adungo Ferdinard, Konongoi Samson Limbaso, Inoue Shingo, Zhan Lin, Sang Rosemary, Ashur Salame, Kwallah Allan Ole, Mwau Matilu, Morita Kouichi, Yu Fuxun
School of Medicine, Guizhou University, Guiyang 550002, China.
Kenya Medical Research Institute, Nairobi 54840-00200, Kenya.
Pathogens. 2024 Jul 13;13(7):582. doi: 10.3390/pathogens13070582.
Rift Valley fever virus (RVFV) belonging to the Phenuiviridae family is responsible for a zoonotic disease called Rift Valley fever (RVF). Currently, RVFV has spread from Africa to Asia, and due to its ability to cause high mortality rates, it has significantly impacted human health and economic development in many societies. Highly specific and sensitive systems for sero-diagnosis of RVFV infection are needed for clinical use.
BALB/c mice were immunized with recombinant RVFV nucleocapsid (rRVFV-N) protein and the spleen cells fused with SP2/0 myeloma cells to create hybridoma cell lines. The secreted monoclonal antibodies (MAbs) were purified and characterized. Enzyme-linked immunosorbent assay (ELISA) systems for the detection of IgG and IgM using the new MAbs were established and evaluated. Serum samples from 96 volunteers and 93 patients of suspected RVF from Kenya were tested compared with the ELISA systems based on inactivated viruses and the rabbit polyclonal antibody.
Three monoclonal antibodies against rRVFV-N protein were established. The performance of the MAb-based sandwich IgG ELISA and the IgM capture ELISA perfectly matched the ELISA systems using the inactivated virus or the polyclonal antibody.
Recombinant RVFV-N protein-specific MAbs were developed and they offer useful tools for RVFV studies. The MAb-based ELISA systems for detecting IgG and IgM offer safe and useful options for diagnosing RVFV infections in humans.
裂谷热病毒(RVFV)属于白蛉病毒科,可引发一种名为裂谷热(RVF)的人畜共患病。目前,RVFV已从非洲传播至亚洲,因其能导致高死亡率,对许多社会的人类健康和经济发展造成了重大影响。临床应用需要高度特异且灵敏的RVFV感染血清诊断系统。
用重组RVFV核衣壳(rRVFV-N)蛋白免疫BALB/c小鼠,其脾细胞与SP2/0骨髓瘤细胞融合以建立杂交瘤细胞系。对分泌的单克隆抗体(MAb)进行纯化和鉴定。建立并评估了使用新MAb检测IgG和IgM的酶联免疫吸附测定(ELISA)系统。将来自96名志愿者和93名肯尼亚疑似裂谷热患者的血清样本与基于灭活病毒和兔多克隆抗体的ELISA系统进行比较检测。
建立了三种针对rRVFV-N蛋白的单克隆抗体。基于MAb的夹心IgG ELISA和IgM捕获ELISA的性能与使用灭活病毒或多克隆抗体的ELISA系统完全匹配。
开发了重组RVFV-N蛋白特异性MAb,它们为RVFV研究提供了有用的工具。基于MAb的检测IgG和IgM的ELISA系统为诊断人类RVFV感染提供了安全且有用的选择。