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目的探讨7种细胞因子的mRNA表达水平作为诊断结核分枝杆菌潜伏感染生物标志物的可能性。方法健康对照64人,活动性肺结核50人,结核分枝杆菌潜伏感染60人,采用实时荧光定量PCR(qRT-PCR)检测结核特异性抗原刺激的外周血中TNF-α、IFN-γ、IL-2、IL-10、IFI35、CXCL10及Foxp3的mRNA表达水平。结果 7种因子相对表达量在活动性肺结核组分别为2.33±0.09、4.94±0.45、150±17.82、2.02±0.55、9.53±5.96、5.56±0.97、1.24±0.17,潜伏感染组分别为3.2±0.61、10.04±1.89、202±17.6、21.89±10.25、29.17±11.19、41.35±11.46、2.14±0.67,健康组分别为1.2±0.08、2.92±0.03、53±17.82、1.48±0.17、1.28±0.77、1.63±0.25、1.03±0.67。潜伏感染组与健康组相比7种细胞因子的mRNA表达水平,差异均有统计学意义(P<0.01),相应细胞因子检测的敏感性、特异性分别为95.1%、82.5%,92.5%、88.7%,91.9%、90.1%,88.2%、51.5%,74.2%、61.2%,81.8%、73.4%和80.4%、51.3%;潜伏感染组和活动性结核组比较除IFI35因子外,其余6种细胞因子的mRNA表达水平差异均有统计学意义(P<0.05),相应细胞因子检测的特异性、敏感性分别为90.6%、80%,88.6%、92.7%,91.9%、86.2%,57.6%、60%,50%、58%,76%、72.2%和66.7%、67.1%;健康对照组和活动性结核组相比较,7种细胞因子的mRNA表达水平差异均有统计学意义(P<0.01),相应细胞因子检测的敏感性、特异性分别为92.3%、90%,88.6%、86.7%,94.4%、86.7%,89.7%、65.8%,82.4%、63.3%,88.2%、70%和92.9%、72.1%。结核病密切接触者潜伏感染率为48.97%,健康组潜伏感染率为36%。结论结核病患者血清TNF-α、IFN-γ、IL-2mRNA水平升高,以潜伏感染者升高更显著,因此可作为诊断结核分枝杆菌潜伏感染的生物标志物。
Objective To explore the mRNA expression levels of seven cytokines as a potential biomarker for the diagnosis of latent infection with Mycobacterium tuberculosis. Methods Sixty-four healthy controls and 50 active pulmonary tuberculosis patients were enrolled in this study. Sixty latent infections of Mycobacterium tuberculosis were detected by real-time quantitative PCR (qRT-PCR). The levels of TNF-α, IFN-γ and IL- -2, IL-10, IFI35, CXCL10 and Foxp3 mRNA expression levels. Results The relative expression levels of seven factors in active tuberculosis group were 2.33 ± 0.09, 4.94 ± 0.45, 150 ± 17.82, 2.02 ± 0.55, 9.53 ± 5.96, 5.56 ± 0.97 and 1.24 ± 0.17, respectively. The latent infection group was 3.2 ± 0.61 , 10.04 ± 1.89, 202 ± 17.6, 21.89 ± 10.25, 29.17 ± 11.19, 41.35 ± 11.46, 2.14 ± 0.67 respectively, and the healthy group was 1.2 ± 0.08, 2.92 ± 0.03, 53 ± 17.82, 1.48 ± 0.17, 1.28 ± 0.77, 1.63 ± 0.25, 1.03 ± 0.67. The sensitivity and specificity of the corresponding cytokines were 95.1%, 82.5%, 92.5%, respectively, compared with those in the latent infection group and the healthy group (P <0.01) 88.7%, 91.9%, 90.1%, 88.2%, 51.5%, 74.2%, 61.2%, 81.8%, 73.4% and 80.4%, 51.3% respectively. In addition to the IFI35 factor, latent infection group and active tuberculosis group were the other 6 The specificity and sensitivity of cytokines were 90.6%, 80%, 88.6%, 92.7%, 91.9%, 86.2%, 57.6% respectively. There was significant difference between the two groups (P <0.05) , 60%, 50%, 58%, 76%, 72.2% and 66.7%, 67.1% respectively. There were significant differences in the mRNA expression levels of the seven cytokines between the healthy control group and the active tuberculosis group (P < 0.01). The sensitivity and specificity of corresponding cytokines were 92.3%, 90%, 88.6%, 86.7%, 94.4%, 86.7%, 89.7%, 65.8%, 82.4%, 63.3%, 88.2%, 70% And 92.9%, 72.1% respectively. The latent infection rate of close contact with tuberculosis was 48.97%, and the latent infection rate of healthy group was 36%. Conclusions The levels of TNF-α, IFN-γ and IL-2 mRNA in tuberculosis patients are elevated, especially in those with latent infection. Therefore, it can be used as a biomarker to diagnose the latent infection of Mycobacterium tuberculosis.