PDF(Pubmed)
Abstract：
BACKGROUND: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
摘要：
背景：属于Phenuiviridae家族的裂谷热病毒（RVFV）是一种称为裂谷热（RVF）的人畜共患疾病的原因。目前,RVFV已经从非洲传播到亚洲,并且由于其导致高死亡率的能力，它严重影响了许多社会的人类健康和经济发展。临床需要用于RVFV感染的血清诊断的高度特异性和敏感性系统。
方法：用重组RVFV核衣壳（rRVFV-N）蛋白免疫BALB/c小鼠，脾细胞与SP2/0骨髓瘤细胞融合，建立杂交瘤细胞系。纯化并表征分泌的单克隆抗体(MAb)。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。与基于灭活病毒和兔多克隆抗体的ELISA系统相比，测试了来自肯尼亚的96名志愿者和93名疑似RVF患者的血清样品。
结果：建立了三种抗rRVFV-N蛋白的单克隆抗体。基于MAb的夹心IgGELISA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。
结论：开发了重组RVFV-N蛋白特异性单克隆抗体，它们为RVFV研究提供了有用的工具。用于检测IgG和IgM的基于MAb的ELISA系统为诊断人的RVFV感染提供了安全和有用的选择。
方法：用重组RVFV核衣壳（rRVFV-N）蛋白免疫BALB/c小鼠，脾细胞与SP2/0骨髓瘤细胞融合，建立杂交瘤细胞系。纯化并表征分泌的单克隆抗体(MAb)。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。与基于灭活病毒和兔多克隆抗体的ELISA系统相比，测试了来自肯尼亚的96名志愿者和93名疑似RVF患者的血清样品。
结果：建立了三种抗rRVFV-N蛋白的单克隆抗体。基于MAb的夹心IgGELISA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。
结论：开发了重组RVFV-N蛋白特异性单克隆抗体，它们为RVFV研究提供了有用的工具。用于检测IgG和IgM的基于MAb的ELISA系统为诊断人的RVFV感染提供了安全和有用的选择。
