结核分枝杆菌底物蛋白rRv1976c在结核病辅助诊断中的价值

Value of Mycobacterium tuberculosis substrate protein rRv1976c in the auxiliary diagnosis of tuberculosis

目的检测并比较结核分枝杆菌底物蛋白rRv1976c刺激活动性结核病患者(ATBs)、潜伏感染者(LTBIs)和健康对照者(HCs)外周血T细胞产生IFN-γ水平,评价rRv1976c在诊断结核病中的潜在价值。方法构建重组载体pPROEX-Rv1976c,表达并纯化底物蛋白rRv1976c;从山西省长治医学院和平医院募集32例受试者,依照rEC-全血IFN-γ分析试验(rEC-WBIA)诊断标准和临床诊断标准分组并进行WBIA试验。结果成功构建pPROEX-Rv1976c质粒,SDS-PAGE鉴定表达产物rRv1976c分子量为15.96 kD,与预期相符,蛋白质印迹法证实表达正确;rRv1976c刺激ATBs外周血T细胞产生的IFN-γ水平显著高于LTBIs(u=3.783,P<0.05)和HCs(u=3.907,P<0.05),LTBIs组也显著高于HCs(u=3.674,P<0.05)。结论rRv1976c在结核病辅助诊断中具有潜在价值。

Objective To detect and compare the levels of IFN-γin peripheral blood T cells stimulated by Mycobacterium tuberculosis substrate protein rRv1976c in patients with active tuberculosis(ATBs),latent TB Infection(LTBIs)and healthy controls(HCs),and to evaluate the potential value of rRv1976c in the auxiliary diagnosis of tuberculosis.Methods The recombinant pPROEX-Rv1976c was constructed,and the substrate protein rRv1976c was expressed and purified.A total of 32 subjects were recruited from Heping Hospital of Changzhi Medical College of Shanxi Province,and rEC-WBIA(rEC-whole blood IFN-γrelease assay)diagnostic criteria and clinical diagnostic criteria were performed for WBIA assay.Results The plasmid of pPROEX-Rv1976c was successfully constructed,and the molecular weight of rRv1976c was 15.96 kD by SDS-PAGE,which was consistent with the expectation and confirmed by Western blotting.The level of IFN-γin ATBs peripheral blood T cells stimulated by rRv1976c was significantly higher than that of LTBIs(u=3.783,P<0.05)and that of HCs(u=3.907,P<0.05),and that of LTBIs group was also significantly higher than that of HCs(u=3.674,P<0.05).Conclusion rRv1976c has potential value in the diagnosis of tuberculosis.