荧光定量PCR在脑脊液结核分枝杆菌DNA检测中的应用

摘要

Objective To use fluorescence quantitation polymerase chain reaction(PCR) for determining Mycobacterium tuberculosis(MTB) DNA in cerebrospinal fluid,and to investigate the clinical significance for the diagnosis of tuberculous meningitis. Methods A total of 108 clinically-diagnosed or suspected tuberculous meningitis patients and 50 non-tuberculous meningitis patients were enrolled,and their cerebrospinal fluid samples were collected and determined by smear,MTB culturing and fluorescence quantitation PCR. Results The positive rates of smear,MTB culturing and fluorescence quantitation PCR were 9.72%(7/72),16.67%(12/72)and 76.39%(55/72) in 72 clinically-diagnosed patients with tuberculous meningitis. The positive rates of smear,MTB culturing and fluorescence quantitation PCR were 5.55%(2/36),11.11%(4/36) and 25.00%(9/36) in 36 suspected patients with tuberculous meningitis. The difference of positive rates among the 3 methods was significant statistically(P<0.05). Cerebrospinal fluid samples of 50 non-tuberculous meningitis patients determined by smear, MTB culturing and fluorescence quantitation PCR were negative. Conclusions Fluorescence quantitation PCR for determining Mycobacterium tuberculosis can be used for the diagnosis and identification of tuberculous meningitis.%目的 应用荧光定量聚合酶链反应(PCR)检测结核性脑膜炎患者脑脊液结核分枝杆菌(MTB)DNA,探讨其对结核性脑膜炎的诊断价值.方法 选取108例临床确诊或疑似的结核性脑膜炎患者和50例非结核性脑膜炎患者脑脊液样本,采用抗酸染色涂片法、MTB培养法及荧光定量PCR进行检测,对不同组间的检测结果 进行对比分析.结果 108例脑膜炎患者中,确诊结核性脑膜炎72例,抗酸染色涂片法、MTB培养法及荧光定量PCR的阳性率分别为9.72%(7/72)、16.67%(12/72)及76.39%(55/72);疑似结核性脑膜炎36例,抗酸染色涂片法、MTB培养法及荧光定量PCR的阳性率分别为5.55%(2/36)、11.11%(4/36)及25.00%(9/36),2个组间阳性率差异有统计学意义(P<0.05).50例非结核性脑膜炎患者脑脊液样本抗酸染色涂片法、MTB培养法及荧光定量PCR检测均为阴性.结论 荧光定量PCR检测脑脊液中的MTB DNA对诊断和鉴别诊断结核性脑膜炎具有较高的临床价值.