新型布尼亚病毒抗体间接ELISA检测方法的建立  

Establishment of a novel bunyavirus IgG indirect ELISA method

作　　者：李欣昱 谭凌云 孙千姿 李德雨 赵红[3] 赵怀龙[3] 杨盼 韩越 郑学星 贺威 王慧 赵忠鹏 LI Xinyu;TAN Lingyun;SUN Qianzi;LI Deyu;ZHAO Hong;ZHAO Huailong;YANG Pan;HAN Yue;ZHENG Xuexing;HE Wei;WANG Hui;ZHAO Zhongpeng(Department of Immunology,School of Basic Medical Sciences,Anhui Medical University,Hefei 230032,China;State Key Laboratory of Pathogen and Biosecurity,Beijing Institute of Microbiology and Epidemiology,Beijing 100071,China;Institute of Viral Diseases,Jinan Center for Disease Control and Prevention,Jinan 250021,China;School of Public Health,Cheeloo College of Medicine,Shandong University,Jinan 250012,China)

机构地区：[1]安徽医科大学基础医学院,合肥230032 [2]军事科学院军事医学研究院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071 [3]济南市疾病预防控制中心病毒性疾病检验所,250021 [4]山东大学齐鲁医学院公共卫生学院,济南250012

出　　处：《免疫学杂志》2023年第6期539-545,共7页Immunological Journal

基　　金：国家重点研发计划(2021YFD1800500、2022YFC2305000)。

摘　　要：目的本研究旨在建立新型布尼亚病毒抗体检测的高特异性、低成本间接ELISA方法。方法通过优化抗原表达、纯化、包被、血清稀释、孵育时间等条件,以Gn的截短蛋白Gn2为包被抗原,开发了一种间接ELISA方法。检测该方法的稳定性、敏感性、特异性、符合率。结果批内和批间变异系数均小于10%,阳性血清临床符合率达76.9%(10/13),阴性血清临床符合率达86.7%(26/30)。结论本研究所建立的间接ELISA方法可用于临床血清样品的检测,为发热伴血小板减少综合征的疾病诊断和监测提供了较为可靠和廉价的检测方法。Severe fever with thrombocytopenia syndrome(SFTS)that induced by a novel bunyavirus is an emerging zoonotic infectious disease with a mortality rate of 10%-20%.This study was designed to establish a highly specific,low-cost SFTSV IgG indirect ELISA method by optimizing the expression,purification and encapsulation of Gn2 antigen,the dilution of human sera and HRP-conjugated antibody and the incubation time.The stability,sensitivity,specificity and compliance rates of the method were tested.Data showed that the intra-and inter-batch coefficients of variation of the constructed method were less than 10%,and the clinical compliance rate of the IgG assay kit test reached 76.9%(10/13)for positive sera and 86.7%(26/30)for negative sera.The results indicate that the indirect ELISA established in this study can be used for the detection of clinical sera samples and provides a more reliable and low-cost method that meets the needs of disease monitoring.

关 键 词：新型布尼亚病毒 原核表达 Gn2蛋白 间接ELISA 

分 类 号：R373.23[医药卫生—病原生物学]