CAJ | 학술논문

目的探讨重组华支睾吸虫半胱氨酸蛋白酶在华支睾吸虫病血清学诊断上的应用价值. 方法以成虫cDNA为模板PCR扩增GenBank中一个华支睾吸虫半胱氨酸蛋白酶(No. AF093242,简称CysB)的部分基因片段,亚克隆到pTrcHis表达载体,转化入大肠埃希菌DH5α.表达出的重组蛋白经亲合层析纯化后用于ELISA检测华支睾吸虫及其他寄生虫感染血清,评价其诊断应用价值. 结果成功克隆与表达CysB基因片段,纯化后的重组蛋白用于华支睾吸虫病诊断的敏感性达96.0%(48/50),与其他寄生虫病的总交叉反应为3.8%(1/26),而对比实验中可溶性粗抗原(CAA)用于检测的敏感性为88.0%(44/50),与其他寄生虫感染血清无交叉反应. 结论重组华支睾吸虫半胱氨酸蛋白酶(CysB)用于ELISA诊断技术具有较高的敏感性及特异性,有希望成为可溶性粗抗原的替代之一.
목적탐토중조화지고흡충반광안산단백매재화지고흡충병혈청학진단상적응용개치. 방법이성충cDNA위모판PCR확증GenBank중일개화지고흡충반광안산단백매(No. AF093242,간칭CysB)적부분기인편단,아극륭도pTrcHis표체재체,전화입대장애희균DH5α.표체출적중조단백경친합층석순화후용우ELISA검측화지고흡충급기타기생충감염혈청,평개기진단응용개치. 결과성공극륭여표체CysB기인편단,순화후적중조단백용우화지고흡충병진단적민감성체96.0%(48/50),여기타기생충병적총교차반응위3.8%(1/26),이대비실험중가용성조항원(CAA)용우검측적민감성위88.0%(44/50),여기타기생충감염혈청무교차반응. 결론중조화지고흡충반광안산단백매(CysB)용우ELISA진단기술구유교고적민감성급특이성,유희망성위가용성조항원적체대지일.
Objective To evaluate the value of a recombinant Clonorchis sinensis cysteine proteinase for the serological diagnosis of clonorchiasis. Methods Based on the sequence (GenBank, No. AF093242), a cysteine proteinase (CysB) gene fragment of C. sinensis was cloned into pTrcHis expression vector and transformed into an Escherichia coli DH5α strain. The recombinant proteins expressed in the bacteria were purified by affinity chromatography and then probed by enzymed-linked immunosorbent assay (ELISA) with sera from patients with clonorchiasis and other parasitic infections. Results For the tested sera, ELISA showed that CysB, judged by a calculated cut-off value, had a sensitivity of 96.0% (48/50) for the diagnosis of clonorchiasis and a cross-reactivity of about 3.8% (1/26) with other parasitic diseases, while C. sinensis adult worm antigen (CAA) had a sensitivity of 88.0% and no cross-reaction was found. Conclusion The results suggested that the recombinant C. sinensis CysB, although having weak cross-reaction with some cases of other parasitic diseases, was valuable for the diagnosis of clonorchiasis and was promising to be developed as an alternative diagnostic antigen.