Rv2031c、38kDa及融合蛋白CFP10-ESAT6用于结核病血清学诊断的评价

发布时间：2018-08-22 15:10

【摘要】：目的对Rv2031c、38kDa以及融合蛋白CFP10-ESAT6三种结核分枝杆菌抗原和抗原的混合物进行检测效能评价,筛选最佳抗原并分析其应用价值。方法以结核分枝杆菌标准株H37Rv菌株基因组为模板,PCR扩增目的基因;利用pET-30a、pET-32a载体构建重组质粒,在宿主E.coli BL21(DE3)细胞中诱导表达,利用His标签亲和镍柱纯化重组蛋白。分别用单种重组蛋白和几种蛋白混合后包被酶标板,采用ELISA对129份血清(39份结核病患者血清和90份健康者血清)进行检测。结合受试者工作特征曲线法计算不同抗原和混合抗原的最佳临界值(Cut-off value)包被浓度,得出检测的灵敏度、特异度和曲线下面积,对其诊断效能和应用价值进行综合评价。结果以Rv2031c、38kDa和CFP10-ESAT6为包被抗原的ELISA法检测相应抗体的灵敏度分别为41.0%,51.3%和46.2%,特异度分别为90.0%、88.9%和79.1%,曲线下面积分别为0.640、0.761和0.690,以3种蛋白混合物为包被抗原的3项指标分别为66.7%、77.8%和0.746。结论 Rv2031c、38kDa以及融合蛋白CFP10-ESAT6 3种抗原混合物的诊断效能较好,具有一定的临床应用价值。
[Abstract]:Objective to evaluate the efficiency of detecting the mixture of Rv2031cP38kDa and fusion protein CFP10-ESAT6, and to screen the best antigen and analyze its application value. Methods the target gene was amplified from the genome of Mycobacterium tuberculosis standard strain H37Rv, and the recombinant plasmid was constructed by pET-30a pET-32a vector. The recombinant protein was induced and expressed in host E.coli BL21 (DE3) cells. The recombinant protein was purified by His tag affinity nickel column. ELISA was used to detect 129 sera (39 sera from tuberculosis patients and 90 sera from healthy people). The optimal critical value (Cut-off value) of different antigens and mixed antigens (Cut-off value) was calculated by using the method of operating characteristic curve. The sensitivity, specificity and area under the curve were obtained. The diagnostic efficacy and application value were evaluated synthetically. Results the sensitivities of Rv2031cn38kDa and CFP10-ESAT6 for the detection of the corresponding antibodies were 41. 0% and 46. 2%, the specificity was 90. 088. 9% and 79. 1%, respectively. The areas under the curve were 0. 640, 0. 761 and 0. 690, respectively. The three indexes of the three protein mixtures as coated antigens were 67. 8% and 0. 746, respectively. Conclusion Rv2031 ct 38kDa and fusion protein CFP10-ESAT6 have good diagnostic efficacy and have certain clinical application value.
【作者单位】：北京市朝阳区疾病预防控制中心;传染病预防控制国家重点实验室中国疾病预防控制中心传染病预防控制所/传染病预防控制国家重点实验室;
【基金】：国家"艾滋病和病毒性肝炎等重大传染病防治科技重大专项"课题(No.2013ZX10003006)资助
【分类号】：R52

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