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
孙永鹏1,靳输梅1,詹炉停1,温桂平1,2,赵 敏1,2,张 伟1,2,夏宁邵1,2,郑子峥1*

(1.厦门大学分子疫苗学与分子诊断学国家重点实验室,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)

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

Double-antibody Sandwich ELISA for Detecting Respiratory Syncytial Virus in Clinical Nasopharynx Aspirate Samples
SUN Yongpeng1,JIN Shumei1,ZHAN Luting1,WEN Guiping1,2,

(1.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics,Xiamen University,2.National Institute of Diagnostics and Vaccine Development of Infectious Diseases,School of Public Health,Xiamen University,Xiamen 361102,China)

respiratory syncytial virus(RSV); fusion protein; double-antibody sandwich ELISA; rapid diagnosis

DOI: 10.6043/j.issn.0438-0479.201607004

备注

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.