中国动物检疫Issue(12)：65-69,5.
猪流行性腹泻病毒双抗夹心ELISA的建立
Development of a Double Antiby Sandwich ELISA for Detection of Porcine Epidemic Diarrhea Virus
摘要
Abstract
Porcine epidemic diarrhea virus(PEDV)was used to immunize laying hens for preparation of speciifc yolk antibody(IgY)as capturing antibody. A double antibody sandwich ELISA(DAS-ELISA)was developed for the detection of PEDV in pigs. The optimal coating concentration of anti-PEDV IgY was 1:4000(12.35µg/mL);the opti-mal concentration of mouse-anti-PEDV polyclonal antibody was 1:6000(10.25µg/mL);optimal reaction time was 30 minutes;optimal working concentration of HRP-labelled goat-anti-mouse IgG was 1:6000;and the positive standard value was OD450≥0.130. The DAS-ELISA showed no cross-reaction with porcine transmissible gastroenteritis virus, porcine rotavirus,Escherichia coli k88/k99/987P/F41. One hundred clinical fecal samples which had been detected by real-time PCR were analyzed by this method. 6 out of 8 real-time PCR positive samples were positive in the DAS-ELISA,and all the negative samples in real-time PCR were negative in the DAS-ELISA. The results indicated that the DAS-ELISA was highly speciifc,reproducible and sensitive,and suitable for rapid detection of PEDV.关键词
猪流行性腹泻病毒/卵黄抗体/双抗夹心ELISAKey words
porcine epidemic diarrhea virus(PEDV)/IgY/double antibody sandwich ELIS分类
农业科技引用本文复制引用
雷利,彭姣,祁振强,秦智峰,卢超..猪流行性腹泻病毒双抗夹心ELISA的建立[J].中国动物检疫,2014,(12):65-69,5.基金项目
深圳市科创委技术创新计划专项基金资助项目， No. CXZZ20130320154210829 （）
