Kim Sunghyun, Lee Hyejon, Kim Hyunjung, Kim Yeun, Cho Jang-Eun, Jin Hyunwoo, Kim Dae Yeon, Ha Sang-Jun, Kang Young Ae, Cho Sang-Nae, Lee Hyeyoung
Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Republic of Korea; Institute for Life Science and Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul, Republic of Korea.
Department of Microbiology, Yonsei University College of Medicine, Seoul, Republic of Korea.
J Mol Diagn. 2015 Jan;17(1):90-9. doi: 10.1016/j.jmoldx.2014.08.005.
Interferon (IFN)-γ release assays have limited sensitivity and cannot differentiate between active tuberculosis (TB) disease and latent TB infection (LTBI). Numerous cytokines and regulator factors have been implicated in the pathogenesis and control of Mycobacterium tuberculosis infection. Additional cytokines and chemokines associated with M. tuberculosis infection may improve the performance of IFN-γ release assays. We developed a real-time RT-PCR TaqMan assay for targeting levels of eight human targets [IFN-γ, tumor necrosis factor (TNF)-α, IL-2R, IL-4, IL-10, CXCL9, CXCL10, and CXCL11] and evaluated the assay with three different study groups. Results showed that the sensitivity of TNF-α, IL-2R, and CXCL10 in the active pulmonary tuberculosis (PTB) group was 96.43%, 96.43%, and 100%, respectively. The sensitivity of IL-2R and CXCL10 in the latent tuberculosis infection group was 86.36% and 81.82%, respectively. Statistical results showed that TNF-α and CXCL9 were the best individual markers for differentiating between the PTB, LTBI, and non-TB groups. For optimal sensitivity and differentiation of M. tuberculosis infection status, the simultaneous detection of multiple targets was attempted. The combination of IFN-γ, TNF-α, and IL-2R, and the combination of TNF-α, IL-2R, CXCL9, and CXCL10 showed the best performance for detecting active PTB (both 100% positivity) and LTBI (86.36% and 81.82% positivity, respectively). These results imply that the combination of suitable markers is useful in efficiently diagnosing TB and differentiating M. tuberculosis infection status.
干扰素（IFN）-γ释放试验敏感性有限，无法区分活动性结核病（TB）和潜伏性结核感染（LTBI）。众多细胞因子和调节因子与结核分枝杆菌感染的发病机制及控制有关。与结核分枝杆菌感染相关的其他细胞因子和趋化因子可能会提高IFN-γ释放试验的性能。我们开发了一种实时逆转录聚合酶链反应（RT-PCR）TaqMan检测方法，用于检测8种人类靶标[IFN-γ、肿瘤坏死因子（TNF）-α、白细胞介素-2受体（IL-2R）、白细胞介素-4、白细胞介素-10、CXC趋化因子配体9（CXCL9）、CXC趋化因子配体10（CXCL10）和CXC趋化因子配体11（CXCL11）]的水平，并在三个不同研究组中对该检测方法进行了评估。结果显示，活动性肺结核（PTB）组中TNF-α、IL-2R和CXCL10的敏感性分别为96.43%、96.43%和100%。潜伏性结核感染组中IL-2R和CXCL10的敏感性分别为86.36%和81.82%。统计结果表明，TNF-α和CXCL9是区分PTB、LTBI和非TB组的最佳单项标志物。为了实现对结核分枝杆菌感染状态的最佳敏感性和区分能力，我们尝试同时检测多个靶标。IFN-γ、TNF-α和IL-2R的组合，以及TNF-α、IL-2R、CXCL9和CXCL10的组合在检测活动性PTB（阳性率均为100%）和LTBI（阳性率分别为86.36%和81.82%）方面表现最佳。这些结果表明，合适标志物的组合有助于有效诊断结核病并区分结核分枝杆菌感染状态。
