西尼罗病毒抗体酶联免疫吸附试验检测方法的评价  

Evaluation of enzyme linked immunosorbent assay for the detection of West Nile virus antibodies

作　　者：何虎鹏[1] 张久松[2] 丁国武[1] 张泮河[2] 刘一萍[2] 高玉然[2] 曹务春[2] 

机构地区：[1]兰州大学公共卫生学院,兰州730000 [2]军事医学科学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

出　　处：《中国人兽共患病学报》2007年第2期121-123,共3页Chinese Journal of Zoonoses

基　　金：国家自然科学基金项目(No.30471495);全军医学科学技术"十一五"计划重大项目(06D006-1)

摘　　要：目的系统地评价血清西尼罗病毒(West Nile virus,WNV)抗体的酶联免疫吸附试验(ELISA)检测方法,为在人群和宿主动物中进行WNV感染血清流行病学调查提供技术支持。方法利用构建的包膜蛋白重组质粒(pQE-30)表达纯化西尼罗病毒包膜蛋白,以此为抗原进行ELISA检测。在对抗原包被量、酶标抗体浓度、血清稀释度进行优化的基础上,对本方法的灵敏度和特异度进行评价。结果确定最适抗原包被量为0.034μg,酶标抗体工作浓度和血清稀释度分别为1∶4 800和1∶80;批内变异和批间变异分别为6.7%和22.6%;对小鼠WNV抗体阳性血清53份、JEV抗体阳性血清48份和阴性对照血清94份进行检测,灵敏度为86.8%,特异度分别为93.8%和92.6%。结论本研究建立的ELISA方法灵敏、检测抗体特异,结果可重复,是一种有价值的血清学调查方法。In order to provide technical support for the sero-epidemiologic survey in human population and animal hosts, the enzyme linked immunosorbent assay（ELISA） for the detection of West Nile virus（WNV） antibodies in sera was established and evaluated. Using the recombinant envelope protein of WNV as antigen, the serum antibody was detected by ELISA. After the parameters of the assay were optimized, the reproducibility and the sensitivity and specificity of the method were evaluated. The optimal coating amount of antigen was 0. 034μg. and the dilutions of enzyme labelled antibody and serum were 1 ： 4 800 and 1 ： 80, respectively. While the intra and inter coefficient variations（CV） were 6.7% and 22.6% respectively. The sensitivity and specificity of this method was 86.8% and 92. 6% in the detections of 53 serum samples with WNV antictodies and 94 negative control sera, and the specificity was 93. 8% in the detections of 48 serum samples with Japanese encephalitis virus （JEV） antibodies. In conclusion, the ELISA established in this study has higher sensitivity and specificity,and was valuable for sero-epidemiologic survey of WNV infection.

关 键 词：西尼罗病毒 包膜蛋白 酶联免疫吸附试验 

分 类 号：R373[医药卫生—病原生物学]