猪沙波病毒抗体间接ELISA检测方法的建立及初步应用  

Establishment and primary application of an indirect ELISA for porcine sapovirus

作　　者：蒋静[1] 王艳[1] 华修国[2] 陈琰[2] 沈权[2] 

机构地区：[1]上海出入境检验检疫局,上海200135 [2]上海交通大学农业与生物学院,上海200240

出　　处：《畜牧与兽医》2012年第3期13-16,共4页Animal Husbandry & Veterinary Medicine

摘　　要：以原核表达重组蛋白作为抗原建立了猪沙波病毒(SaV)抗体的间接ELISA检测方法。使用矩阵滴定法确定抗原包被浓度为6.4μg/mL,血清稀释倍数为1∶50。试验选用1%BSA作为封闭液,血清和二抗的最佳作用时间分别为60 min和40 min。选择无致癌的TMB为底物,确定TMB最佳反应时间为10 min。通过对20份猪阴性血清样品的检测,计算阳性判定值为0.203。通过交叉试验、批内和批间重复性试验证明,本研究建立的SaV间接ELISA检测方法具有特异性高,重复性好的特点,可用于猪沙波病毒抗体的检测。An indirect ELISA was established for detecting porcine sapovirus using the recombinant protein as antigen. The working con- centration of the antigen was 6. 4 μg/mL and the dilution was 1 ： 50 as determined by matrix titration. 1% BSA was used as blocking solu- tion. The optimized reaction times of serum and anti-porcine IgG were 60 min and 40 min, respectively. The substrate was non-carcinogen TMB and the best reaction time was 10 min. The positive value was 0. 203, which was determined on the basis of the detection results for 20 negative porcine sera. The across reaction test and within-batch and between-batch reproducibility test demonstrated that, the indirect ELISA method established in the study has a high specialty and good reproducibility, and can be applied for the serological detection of porcine sapo- virus.

关 键 词：猪沙波病毒 间接ELISA 抗体 

分 类 号：S854.43[农业科学—临床兽医学]