犬细粒棘球绦虫血清诊断重组epc1蛋白的克隆与表达

S. Kordafshari, S. Hosseini, Hagos Gebrekidan, F. Jalousian, P. Shayan, M. Bandehpour, B. Kazemi, M. Rajabibazl, Malcolm K. Jones, S. Jafariaskari
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引用次数: 0

摘要

细粒棘球绦虫被认为是一种重要的寄生虫感染,分布在世界各地。本研究的目的是在重组系统中表达诊断抗原EpC1,并建立一种间接ELISA诊断感染犬的方法。从屠宰场自然感染的羊的肝脏和肺中采集包虫囊,从原头节中合成cDNA。以pET28a为表达载体,通过His-tag亲和层析纯化,在大肠杆菌中以重组蛋白形式表达EpC1。重组EpC1的表达通过SDS-PAGE和T7-Tag单克隆抗体和自然感染和实验攻毒犬的血清进行Western免疫印迹检测。采用间接ELISA法评估重组EpC1对22种犬血清的诊断潜力。ELISA检测的敏感性为100%,特异性为93.3%,阳性预测值为88.8%,阴性预测值为100%。重组EpC1在犬棘球蚴病的诊断中显示出良好的结果。
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Molecular cloning and expression of a recombinant epc1 protein for serodiagnosis of echinococcus granulosus in dogs
Echinococcus granulosus is considered as a significant parasitic infection with worldwide distribution. The present study was aimed to express the diagnostic antigen EpC1 in a recombinant system and set up an indirect ELISA for diagnosis of infected canines. Hydatid cysts were collected from liver and lungs of naturally infected sheep from slaughter houses and cDNA was synthesized from the protoscoleces. The EpC1 was expressed as recombinant protein in E. coli using pET28a as expression vector and purified by His-tag affinity chromatography. Expression of recombinant EpC1 was confirmed using SDS-PAGE followed by Western immunoblotting using T7-Tag monoclonal antibody and sera from naturally infected and experimentally challenged dogs. The diagnostic potential of the recombinant EpC1 was assessed against 22 dog sera using indirect ELISA. The sensitivity, specificity, positive and negative prediction values of the ELISA test were calculated as 100%, 93.3%, 88.8% and 100%, respectively. Recombinant EpC1 showed promising results for the diagnosis of echinococcosis in dogs.
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来源期刊
Journal of Veterinary parasitology
Journal of Veterinary parasitology Veterinary-Veterinary (all)
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