基于微小隐孢子虫重组CP15/60蛋白的间接ELISA检测方法的建立及上海市猪隐孢子虫感染情况调查

DEVELOPMENT OF INDIRECT ELISA WITH RECOMBINANT CP15/60 OF CRYPTOSPORIDIUM PARVUM AND SEROLOGICAL SURVEY OF PIG CRYPTOSPORIDIOSIS IN SHANGHAI

为建立敏感、特异的猪隐孢子虫感染血清学检测方法,了解上海市猪隐孢子虫感染情况,以纯化的重组CP15/60(r CP15/60)为诊断抗原,建立了检测隐孢子虫感染的间接ELISA方法,并对上海市猪隐孢子虫感染情况进行血清学调查。结果显示,与Nested PCR方法检测结果相比,本研究建立的r CP15/60-ELISA方法的阴阳性符合率为83.3%,敏感性为100%,特异性为80%;重复性试验的变异系数在12.5%之内;对341份猪田间血清进行检测,阳性为194份,占56.89%,表明该方法可用于实验室初步诊断和现场猪隐孢子虫病的流行病学调查。

To develop sensitive and specific serological detection method of pig Cryptosporidium infection and understand the situation of Cryptosporidium infection in pigs in Shanghai, an indirect ELISA method were developed to detect Cryptosporidium infection in pigs with the recombinant CP15/60（rCP15/60） protein as diagnostic antigen. The reproducibility, sensitivity and specificity of the method were evaluated. The situation of pigs Cryptosporidium infection were serological detected with the developed method. The result showed that comparing with Nested PCR detection results, the negative and positive coincidence, sensitivity and specificity were 83.3%, 100% and 80%, respectively. The coefficient of variability of reproducibility was≤12.5%. Total of 194（56.89%） sera were positive in 341 sera collected from pigs in field in Shanghai. These results indicated that the method was suitable to be used as laboratory primary diagnosis and field serological survey of porcine cryptosporidiosis.