新孢子虫和弓形虫ELISA及western blot检测方法的建立及应用

为建立牛新孢子虫和弓形虫的免疫学检测方法,并调查新疆部分地区牛新孢子虫和弓形虫的感染情况,本研究应用纯化的新孢子虫重组蛋白SRS2(NcSRS2)和弓形虫重组蛋白SAG2(TgSAG2)作为包被抗原,分别建立新孢子虫和弓形虫的ELISA和western blot血清学检测方法,并进行特异性和重复性试验,以其检测662份疑似样品,并与商品化试剂盒检测结果比较验证。特异性和重复性试验结果表明,建立的方法特异性强、重复性良好。采用建立的两种ELISA方法对662份临床样品的检测结果表明,新孢子虫和弓形虫的抗体阳性率分别为13.44%(89/662)和5.29%(35/662);与IDEXX试剂盒和永辉试剂盒的符合率分别为94.11%和95.92%。此外,western blot检测的新孢子虫和弓形虫抗体阳性率分别为5.14%(34/662)和3.17%(21/662);与建立的ELISA检测方法的符合率分别为91.69%和97.89%。本研究为分析奶牛流产的原因提供了一定的依据。

Establishment and application of indirect ELISA assays and western blot to detect antibodies agaist Neospora caninum and Toxoplasma gondii

Neospora caninum and Toxoplasma gondii are closely related protozoan parasites which causes of abortion and congenital disease in ruminants.To establish the methods for detection of the antibodies against N.caninum and T.gondii in bovine,The indirect ELISAs and western blot assay were developed based on purified recombinant protein SRS2 of N.caninum and SAG2 of T.gondii,respectively.The results show that these methods were highly specificity and repeatability.A total of 662 bovine serum samples were tested and the positive rates for N.caninum and T.gondii were 13.44%(89/662) and 5.29%(35/662) detected by the developed ELISAs,which were 94.11% and 95.92% agreements with ’IDEXX’ N.caninum Antibody Test Kit and ’Yong hui’ T.gondii Antibody Test Kit,respectively.In addition,the positive rates for N.caninum and T.gondii were 5.14%(34/662) and 3.17%(21/662) using western blot,and the agreements with the developed ELISAs were 91.69% and 97.89%,respectively.This study provides basis for interpretation of the bovine abortion.