基于GAPDH重组蛋白建立羊肝片吸虫病间接ELISA检测方法  

Establishment of an Indirect ELISA Based on GAPDH Recombinant Protein for Diagnosis of Ovine Fascioliasis

作　　者：孙益春 魏雅婷 郑芦珊 杨莹 张金鹏 庞紫璇 路义鑫[1] SUN Yi-chun;WEI Ya-ting;ZHENG Lu-shan;YANG Ying;ZHANG Jin-peng;PANG Zi-xuan;LU Yi-xin(Heilongjiang Provincial Key Laboratory of Zoonosis,College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学动物医学学院黑龙江省动物源性人兽共患病重点实验室,黑龙江哈尔滨150030

出　　处：《中国兽医杂志》2022年第2期34-38,共5页Chinese Journal of Veterinary Medicine

基　　金：国家重点研发计划项目(2017YFD0501200)。

摘　　要：为了建立早期羊肝片吸虫病的血清学快速检测方法,本试验成功克隆并表达肝片吸虫GAPDH重组蛋白,Western blot结果显示,该重组蛋白具有良好的抗原活性。用肝片吸虫GAPDH重组抗原包被,建立肝片吸虫病血清抗体的间接ELISA方法;随后优化反应的最佳血清稀释度和抗原包被量,同时筛选其他反应条件。结果显示,间接ELISA方法批内和批间重复试验的最大变异系数均小于10%,GAPDH重组抗原与华枝睾吸虫、日本血吸虫和捻转血矛线虫阳性血清均无交叉反应。对来自黑龙江省各地区的96份羊血清进行检测,阳性率为16.67%(16/96)。结果表明本试验建立的方法具有良好的敏感性和特异性,能够为早期诊断羊肝片吸虫病提供参考。This study aimed to establish an indirect ELISA for early detection of ovine fascioliasis.The GAPDH gene from Fasciola hepatica was cloned and expressed,and the recombinant GAPDH protein was shown to have good antigen activity as assessed by Western blot.An indirect ELISA was established by using the recombinant GAPDH protein,and serum dilution and quantity of recombinant GAPDH protein required were optimized.The results showed that the indirect ELISA had intra-batch and inter-batch variation of<10%,and no cross reaction between recombinant GAPDH protein and Clonorchis sinensis,Schistosoma japonicum and Haemonchus contortus positive sera.Of the 96 sheep sera from various regions of Heilongjiang province,16(16.67%)were tested positive by the indirect ELISA.Thus,the indirect ELISA provides a sensitive and specific method for the early diagnosis of ovine fascioliasis.

关 键 词：羊 肝片吸虫 GAPDH 间接ELISA 

分 类 号：R383.26[医药卫生—医学寄生虫学]