猪母乳中抗猪传染性胃肠炎病毒IgA检测方法的建立  

Establish the detection method of IgA antibodies against transmissible gastroenteritis virus in sow milk

作　　者：蒋凤英[1] 邹勇[1] 何锡忠[1] 李春华[1] 彭丽英[1] 张婉华[1] 周宗清[1] 张俊平[1] 许宝华[2] 张春玲[1] 徐伟林 

机构地区：[1]上海市农业科学院畜牧兽医研究所,上海201106 [2]南昌大学医学院,江西南昌330006 [3]上海科立特农科(集团)有限公司,上海201106

出　　处：《中国预防兽医学报》2014年第7期551-554,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：江西省自然科学基金项目(2007GZY1003);上海市科委产业化项目(11DZ1930400)

摘　　要：为建立检测猪母乳中抗猪传染性胃肠炎病毒(TGEV)IgA抗体的间接ELISA方法,本研究利用纯化的TGEV作为抗原,通过方阵法确定包被抗原和待检抗体的最佳工作浓度,经过对各种反应条件进行优化,建立了猪母乳抗TGEV IgA抗体检测方法。对TGEV阳性乳汁样品最低检测浓度为1∶320。该方法不与其他常见猪病毒的阳性乳样发生交叉反应,重复性试验的变异系数均小于8.1%。对怀孕母猪免疫后不同时间的乳样检测结果表明该方法与中和试验的符合率达91.5%。本实验建立的检测方法具有良好的敏感性和特异性,为TGEV粘膜疫苗的设计及免疫效果评价提出新的思路。To establish the detection method of IgA antibodies against transmissible gastroenteritis virus （TGEV） from sow milk, the ELISA was developed with purified TGEV as coating antigen. The reaction concentrations of the assay were optimized including： 12 μg/mL of coating antigen, 1% BSA for blocking for 2 hours, 1：40 dilution of milk sample incubating for 60 min, 1：12,000 dilution of rabbit anti-porcine IgA-HRP reacting for 30 min. While, no crossed reactions with other common swine diseases in positive dairy. Variation coefficient of repeatability was less than 8.1%. Detection results from dairy of pregnant sows at different times after immunization indicated that the coincidence rate of this method with neutralization test was 91.5%. The established method for IgA detection in sow milk has high sensitivity and specificity, which proposes new ideas for TGEV vaccine design and immune effect evaluation.

关 键 词：猪传染性胃肠炎病毒 IgA抗体检测 间接ELISA 

分 类 号：S852.65[农业科学—基础兽医学]