中国动物传染病学报2011，Vol.19Issue(2)：43-50,8.
隐孢子虫P23基因的原核表达及间接ELISA检测方法的建立
PROKARYOTIC EXPRESSION OF P23 GENE OF CRYPTOSPORIDIUM MOUSE GENOTYPE AND DEVELOPMENT OF INDIRECT ELISA DETECTION METHOD
摘要
Abstract
The P23 gene of Cryptosporidium mouse genotype was sub-cloned into pGEX-4T-1 vector and expressed in Escherichia coli BL21（DE3）.The recombinant protein rP23 was used to establish an indirect ELISA method for rapid detection of specific Cryptosporidium antibodies in rabbit.The specificity,sensitivity and reproducibility of the method were evaluated,and 23 field rabbit sera samples were detected and compared with faeces detection results of Sheather＇s sucrose flotation and nested PCR.The results showed that the P23 gene was successfully expressed and the recombinant protein rP23 could be recognized by sera from infected rabbits in Western blot test.The indirect ELISA assay（rP23-ELISA） was successfully established using rP23 protein as coating antigen.The results of detection of 23 field sera/faeces samples showed that the rP23-ELISA had higher positive rate comparing with Sheather＇s sucrose flotation and nested PCR,and could be used to further develop an indirect ELISA Kit for the detection of Cryptosporidium antibodies.关键词
隐孢子虫鼠基因型/P23基因/原核表达/间接ELISAKey words
Cryptosporidium mouse genotype/P23 gene/prokaryotic expression/indirect ELISA分类
农业科技引用本文复制引用
张国恩,米荣升,苏庆美,黄燕,周鹏,胡义彬,秦培兰,呼高伟,陈兆国..隐孢子虫P23基因的原核表达及间接ELISA检测方法的建立[J].中国动物传染病学报,2011,19(2):43-50,8.基金项目
国家“十一五”高技术研究发展计划（863)项目（2006AA10A207) （）
国家“十一五”科技支撑计划项目（2007BAD40B05) （）
上海市科技兴农重点攻关项目（沪农科攻字[2005]第3-4号) （）
中央级公益性科研院所基本科研业务费专项资金项目（2010JB12) （）
