犬瘟热病毒(CDV)ELISA检测方法的建立与应用  

Establishment and Application of an ELISA Technique in Detection of Canine Distemper Virus Antibody

作　　者：王吉[1] 卫礼[1] 付瑞[1] 巩薇[1] 岳秉飞[1] 贺争鸣[1] 

机构地区：[1]中国药品生物制品检定所国家实验动物微生物遗传检测中心,北京100050

出　　处：《中国比较医学杂志》2009年第4期59-62,共4页Chinese Journal of Comparative Medicine

摘　　要：目的建立犬CDV抗体ELISA检测方法。方法培养vero细胞,接种CDV病毒,制备vero正常抗原和CDV特异抗原,滴定酶结合物和抗原最佳工作浓度,并进行精密性、敏感性、稳定性、特异性实验。结果正常、特异抗原和酶结合物最佳工作浓度分别为1∶32 000、10μg/mL和1∶8 000;正常、特异抗原批内变异系数分别为9.1%和5.8%,批间平均变异系数分别为8.8%和6.6%;检测灵敏度为1∶2 560;与犬细小病毒(CPV)、犬肝炎病毒(ICHV)均无交叉反应。稳定性试验相对偏差小于25%。结论建立的ELISA方法重复性、稳定性好,特异性、敏感性强。可用于犬CDV抗体的检测。Objective To establishing a method of detection of the canine distemper virus （CDV） antibody with ELISA.Methods Vero cells were cultured and vaccinated with CDV virus to prepare the normal vero antigen and CDV-specific antigen. The best working concentration of enzyme complex and the normal or specific antigen were titrated, and experiments of accuracy, sensitivity, stability and specificity were conducted. Results The best working concentration of normal, specific antigen and the enzyme complex were 1：32000,10 μg/mL and 1：8000, respectively. The inter-assay coefficient of variation of normal antigen and specific antigen was 9.1% and 5.8%, respectively. The mean intra-assay coefficient of variation was 8.8% and 6.6%, respectively. The detection sensitivity was 1：2560. There was no cross-reactivity with dog tiny virus （CPV） and dog hepatitis virus （ICHV）. The stability test showed a relative deviation below 25% . Conclusion The above described ELISA method is good in duplication, stability, specificity and sensitivity. This ELISA technique can be used in detection of dog CDV antibody.

关 键 词：犬瘟热病毒 ELISA 

分 类 号：R33[医药卫生—人体生理学]