CAJ | 학술논문

将隐孢子虫鼠基因型（Cryptosporidium mouse genotype）P23基因亚克隆到pGEX-4T-1载体中,并用大肠杆菌BL21（DE3）为宿主菌诱导表达。以纯化的rP23为诊断抗原,建立隐孢子虫间接ELISA检测方法,观察其敏感性、特异性和重复性,并对临床血清样品进行检测。结果隐孢子虫鼠基因型P23基因在大肠杆菌中获得了高效表达,Western blot检测显示rP23能被隐孢子虫感染兔血清识别。以纯化rP23为诊断抗原,成功地建立了检测兔隐孢子虫病的间接ELISA技术。对23份临床血清检测结果显示,该方法检出率高于Sheather＇s蔗糖漂浮法、套式PCR法。本研究结果为研制兔隐孢子虫病诊断试剂盒打下了基础。
장은포자충서기인형（Cryptosporidium mouse genotype）P23기인아극륭도pGEX-4T-1재체중,병용대장간균BL21（DE3）위숙주균유도표체。이순화적rP23위진단항원,건립은포자충간접ELISA검측방법,관찰기민감성、특이성화중복성,병대림상혈청양품진행검측。결과은포자충서기인형P23기인재대장간균중획득료고효표체,Western blot검측현시rP23능피은포자충감염토혈청식별。이순화rP23위진단항원,성공지건립료검측토은포자충병적간접ELISA기술。대23빈림상혈청검측결과현시,해방법검출솔고우Sheather＇s자당표부법、투식PCR법。본연구결과위연제토은포자충병진단시제합타하료기출。
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.