TaqMan-PCR技术诊断结核病的临床应用研究

目的　探讨TaqMan-聚合酶链反应 (TaqMan-PCR)技术在结核病快速诊断中的临床价值。方法 对 155例活动性肺结核患者的痰和外周血、130例结核性胸膜炎患者的胸腔积液和外周血以及 61例结核性脑膜炎患者的脑脊液和外周血应用TaqMan -PCR检测结核分支杆菌DNA ,痰、胸腔积液和脑脊液进行抗酸染色涂片检查 ,另外 ,痰标本还进行了BACTEC和改良罗氏培养；同期以52例肺癌患者的痰和外周血、50例恶性胸腔积液患者的胸腔积液以及 33例健康人的外周血作为对照进行TaqMan-PCR检测。 结果 155例活动性肺结核患者的痰和外周血、130例结核性胸膜炎患者的胸腔积液和外周血以及 61例结核性脑膜炎患者的脑脊液和外周血TaqMan-PCR的阳性率分别为 49.0%和 51.6%、45.4%和 38.5%以及 50.8%和 42.6% ,痰、胸腔积液和脑脊液TaqMan-PCR的阳性率显著高于抗酸染色涂片以及BACTEC和罗氏培养 (P＜0.05)。TaqMan-PCR检测痰、胸腔积液和外周血的特异性分别为 96.2%、98%和 96.5%。结论 TaqMan-PCR具有较高的敏感性和特异性 ,对结核病的快速诊断具有一定的价值。

Study on clinical application of rapid diagnostic tuberculosis by TaqMan-PCR

Objective To evaluate the clinical value of rapid diagnostic tuberculosis using TaqMan polymerase chain reaction(TaqMan-PCR) technique.Methods It was detected to the specimens of the peripheral blood from 155 patients with active pulmonary tuberculosis, 130 patients with tuberculous pleurisy, 61 patients with tubercular meningitis, 52 patients with lung cancer and 33 healthy adults by TaqMan-PCR, and the specimens of pleural effusion from 130 patients with tuberculous pleurisy and 50 patients with malignant pleurisy, the specimens of cerebrospinal fluid from 61 patients with tubercular meningitis and the specimens of sputum from patients with 155 patients with active pulmonary tuberculosis and 52 patients with lung cancer by TaqMan-PCR and smear respectively. The specimens of sputum from patients with active pulmonary tuberculosis were detected simultaneously by culture with BACTEC and Lowenstein-Jensen. Results The positive rates of sputum and peripheral blood from patients with active pulmonary tuberculosis and of pleural effusion and operipheral blood from patients with tuberculous pleurisy and of cerebrospinal fluid and peripheral blood from 61 patients with tubercular meningitis detected by TaqMan-PCR were 49%, 51.6%, 45.4%, 38.5%, 51%and 42.6%, respectively. The positive rates detected by TaqMan-PCR were higher significantly than those detected by smear, culture with BACTEC and Lowenstein-Jensen(P<0.05). The specificity of TaqMan-PCR detecting for sputum, pleural effusion and peripheral blood were 96.2%, 98% and 96.5%.Conclusion TaqMan-PCR method showed higher sensitivity and specificity. It is a useful tool for rapid diagnosis of tuberculosis.