酶联免疫吸附分析双抗夹心法检测临床鼻咽抽吸物中呼吸道合胞病毒  

Double-antibody Sandwich ELISA for Detecting Respiratory Syncytial Virus in Clinical Nasopharynx Aspirate Samples

作　　者：孙永鹏[1] 靳输梅 詹炉停 温桂平[1,2] 赵敏[1,2] 张伟[1,2] 夏宁邵[1,2] 郑子峥[1] 

机构地区：[1]厦门大学分子疫苗学与分子诊断学国家重点实验室,福建厦门361102 [2]厦门大学公共卫生学院,国家传染病诊断试剂与疫苗工程技术研究中心,福建厦门361102

出　　处：《厦门大学学报（自然科学版）》2017年第4期505-510,共6页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金(81401668);NSFC-NIH生物医学合作研究项目(812111615);福建省科技创新平台建设计划项目(2014Y2101)

摘　　要：呼吸道合胞病毒(respiratory syncytial virus,RSV)是引起5周岁以下儿童下呼吸道感染的主要病原体.目前常用的试剂盒是美国Diagnostic Hybrids公司研发的7项呼吸道病毒检测试剂盒,但其价格昂贵,且需荧光显微镜辅助诊断,不利于在基层医疗机构中推广,国内尚无上市的快速诊断试剂用于临床检测.为此建立用于RSV抗原检测的早期快速诊断试剂,并以反转录聚合酶链式反应(RT-PCR)检测为参比,同时使用酶联免疫吸附分析(ELISA)双抗夹心法和免疫荧光法对112份临床鼻咽抽吸物(NPA)样本进行检测.ELISA双抗夹心法和免疫荧光法的灵敏度分别为79.31%和74.14%,特异度分别为100%和96.30%,Kappa值分别为0.787 1和0.689 5,两者均与RT-PCR检测结果具有高度一致性.此外,将ELISA双抗夹心法与免疫荧光法比较,两者无显著差异(p>0.05).综上,ELISA双抗夹心法操作简便,特异性高,有望代替进口检测试剂盒完成RSV感染的早期快速诊断.Respiratory syncytial virus（RSV）is the major cause of hospitalization and medically attended lower respiratory tract infection in children under 5 years old and infects all children by the age of 2.Currently,RSV infected patients are usually detected with the seven respiratory virus diagnostic kit（immunofluorescence,Diagnostic Hybrids company,USA）,which is expensive and judged subjectively.Nowadays,a rapid diagnostic kit for detection of RSV is not available in China.In this study,we established a sandwich ELISA rapid diagnostic method for antigen detection of RSV antigen at the early stage of infection.112 clinical nasopharyngeal aspirate（NPA）samples were detected by sandwich ELISA and fluorescence microscopy diagnosis,with standard RT-PCR method as reference,and the results show that both methods have high consistency with the RT-PCR method.The sensitivity of the sandwich ELISA and fluorescence microscopy diagnosis was respectively 79.31% and 74.14%,with the specificity of 100% and 96.30%,as well as the Kappa value of 0.787 1 and 0.689 5.There was no statistically significant difference between sandwich ELISA and fluorescence microscopy diagnosis（p〉0.05）.In conclusion,our sandwich ELISA method is simple and highly specific,which is expected to replace imported testing kits for rapid diagnosis of early RSV infection.

关 键 词：呼吸道合胞病毒 融合蛋白 酶联免疫吸附分析双抗夹心法 快速诊断 

分 类 号：R373.9[医药卫生—病原生物学]