Cloning and expression of 21.1-kDa tegumental protein of Clonorchis sinensis and human antibody response to it as a trematode-nematode pan-specific serodiagnosis antigen
Cloning and expression of 21.1-kDa tegumental protein of Clonorchis sinensis and human antibody response to it as a trematode-nematode pan-specific serodiagnosis antigen
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华支睾吸虫 21.1 kDa 皮膜蛋白的克隆和表达及其作为吸虫-线虫泛特异性血清诊断抗原的人抗体反应
DOI：
10.1007/s00436-010-2050-4
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发表时间：
2011-01-01
影响因子：
2
通讯作者：
Hu, Xuchu
中科院分区：
文献类型：
--
作者：
Chen, Jiajia;Xu, Hongxu;Hu, Xuchu
通过生物信息学分析，从华支睾吸虫成虫全长cDNA质粒库中识别出编码21.1kDa外皮蛋白（CsTP21.1）的完整cDNA。重组CsTP 21. 1在大肠杆菌中高效表达，亲和层析纯化，Western blotting鉴定。免疫组化结果显示，CsTP 21.1定位于成虫的被膜。ELISA可检测到华支睾吸虫病患者血清中rCsTP 21.1特异性IgG 1、IgG 2和IgG 4亚类，但其敏感性远低于总IgG。66份华支睾吸虫病患者血清IgG检测的敏感性为100%，特异性为95.5%，敏感性与虫载量无关;对86份肝片吸虫、24份日本血吸虫、31份线虫感染者血清的交叉反应率分别为98.8%、83.3%、93.3%，与弓形虫、裂头蚴无交叉反应。这项研究表明，CsTP 21.1是一种吸虫-线虫泛特异性抗原，在开发人类吸虫和线虫感染的通用免疫诊断试剂盒中具有价值。
A complete cDNA encoding a 21.1-kDa tegumental protein (CsTP21.1) was recognized from Clonorchis sinensis adult full-length cDNA plasmid library by bioinformatics analysis. Recombinant CsTP21.1 was highly expressed in Escherichia coli, purified by affinity chromatography, and identified by Western blotting. Immunohistochemistry demonstrated that CsTP21.1 is localized in the tegument of the adult worm. The rCsTP21.1-specific IgG1, IgG2, and IgG4 subclasses could be detected in the sera of clonorchiasis patients by ELISA, but their sensitivity was much lower than that of total IgG. The sensitivity and specificity of IgG in 66 serum samples of clonorchiasis patients were 100% and 95.5%, and the sensitivity was independent of worm loads; the cross-reaction rates in 86, 24, and 31 serum samples from patients infected with Fasciola hepatica, Schistosoma japonicum, and nematode were 98.8%, 83.3%, 93.3%, respectively, whereas no cross-reactions with Toxoplasma gondii and sparganum. This study demonstrated that CsTP21.1 is a trematode–nematode pan-specific antigen that is valuable in the development of a universal immunodiagnostic kit for human trematode and nematode infections.