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

作　　者：刘宇轩[1] 米荣升[1,2] 黄燕[1,2] 于慧珠[1] 胡盼[1] 詹婷婷[1] 周金林[1,2] 陈兆国[1,2] 

机构地区：[1]中国农业科学院上海兽医研究所农业部动物寄生虫学重点开放实验室农业部动物产品质量安全生物性危害因子风险评估实验室(上海),上海200241 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,扬州225009

出　　处：《中国动物传染病学报》2015年第4期37-43,共7页Chinese Journal of Animal Infectious Diseases

基　　金：国家科技重大专项项目(2012ZX10004220);2015年国家畜禽产品质量安全风险评估项目(GJFP201500803);家畜疫病家畜疫病病原生物学国家重点实验室开放基金课题(SKLVEB2013KFKT017);上海市科技兴农重点攻关项目(沪农科攻字[2005]第1-10号[2005]第3-4号)

摘　　要：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.

关 键 词：微小隐孢子虫 重组CP15/60 间接ELISA 调查 

分 类 号：S852.723[农业科学—基础兽医学]