肝吸虫重组组织蛋白酶B-1蛋白酶作为人类华支睾吸虫病血清学诊断抗原的评估。
Evaluation of liver fluke recombinant cathepsin B-1 protease as a serodiagnostic antigen for human opisthorchiasis.
作者信息
Sripa Jittiyawadee, Brindley Paul J, Sripa Banchob, Loukas Alex, Kaewkes Sasithorn, Laha Thewarach
机构信息
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
出版信息
Parasitol Int. 2012 Mar;61(1):191-5. doi: 10.1016/j.parint.2011.05.009. Epub 2011 Jun 17.
A cathepsin B-like cysteine protease belonging to family C1 is abundantly expressed in the transcriptome and proteome of the carcinogenic liver fluke of humans, Opisthorchis viverrini. This enzyme is present in excretory/secretory (ES) products released by parasites cultured in vitro. This study evaluated the performance of recombinant O. viverrini cathepsin B1 (rOv-CB-1) as an antigen for immunodiagnosis of opisthorchiasis. The full length Ov-CB-1 cDNA was cloned and recombinant protein was produced in catalytically active form in Pichia pastoris. The recombinant Ov-CB-1 (rOv-CB-1) was affinity purified via nickel-NTA chromatography and tested in enzyme-linked immunosorbent assays (ELISA) with human sera from an opisthorchiasis endemic area. Sera from egg-positive O. viverrini infections produced a strong IgG antibody response to rOv-CB-1 both in ELISA and immunoblot analysis. The sensitivity and specificity of the ELISA test was 67% and 81%, respectively. These findings support the feasibility of using recombinant Ov-CB-1 in ELISA for the serodiagnosis of human opisthorchiasis.
一种属于C1家族的组织蛋白酶B样半胱氨酸蛋白酶在人类致癌肝吸虫——华支睾吸虫的转录组和蛋白质组中大量表达。这种酶存在于体外培养寄生虫释放的排泄/分泌（ES）产物中。本研究评估了重组华支睾吸虫组织蛋白酶B1（rOv-CB-1）作为华支睾吸虫病免疫诊断抗原的性能。克隆了全长Ov-CB-1 cDNA，并在毕赤酵母中产生了具有催化活性形式的重组蛋白。重组Ov-CB-1（rOv-CB-1）通过镍-亚氨基三乙酸（NTA）色谱进行亲和纯化，并在酶联免疫吸附测定（ELISA）中用来自华支睾吸虫病流行地区的人血清进行检测。在ELISA和免疫印迹分析中，来自华支睾吸虫卵阳性感染的血清对rOv-CB-1产生了强烈的IgG抗体反应。ELISA检测的敏感性和特异性分别为67%和81%。这些发现支持了在ELISA中使用重组Ov-CB-1进行人类华支睾吸虫病血清诊断的可行性。
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