荧光定量PCR技术检测结核分支杆菌DNA的应用价值

目的评价荧光定量PCR技术检测临床标本中结核分支杆菌的应用价值。方法应用荧光定量PCR法及抗酸染色、结核杆菌培养法，对179例活动性肺结核患者晨痰、123例活动性肺结核患者外周血、34例结核性胸膜炎患者胸水、20例非结核性呼吸系统疾病患者的晨痰标本进行检测。结果179例活动性肺结核患者晨痰、123例活动性肺结核患者外周血、34例结核性胸膜炎患者胸水，涂片抗酸染色阳性率分别为20．67％（37／179）、0．00％、0．00％；细菌培养阳性率分别为41．34％（74／179）、0．00％、2．94％（1／34）；荧光定量PCR技术阳性率分别为64．25％（115／179）、38．21％（47／123）、44．12％（15／34）。3种标本荧光定量PCR技术阳性率高于抗酸染色和结核杆菌培养，经统计学处理发现，差异有统计学意义。用荧光定量PCR技术检测临床标本特异性为95．00％。结论荧光定量PCR技术将PCR扩增、荧光探针杂交及检测一体化，在单一管内完成，具有简便、快速、防污染、敏感性及特异性高等优点，是结核病诊断的有效方法之一。

Clinical value of fluorescent quantitative PCR assay for detection of Mycobacterium tuberculosis in clinical specimens

ObjectiveTo evaluate the clinical value of FQ-PCR assay for detection of Mycobacterium tuberculosis in clinical specimens.MethodsSputum specimens of 179 patients with active pulmonary tuberculosis, peripheral blood specimens of 123 patients with active pulmonary tuberculosis, pleural fluid specimens of 34 patients with tuberculous pleurisy and sputum specimens of 20 patients with other respiratory diseases were tested by smear, culture and FQ-PCR assay.ResultsThe positive rate of sputum, blood, and pleurisy smear was 20.67%,0.00%,0.00% respectively. The positive rate of culture was 41.34%,0.00%, and 2.94% respectively. The positive rate of FQ-PCR assay was 64.25%, 38.21%, and 44.12% respectively. The positive rate of FQ-PCR assay was higher than that of smear and culture significantly (P<0.001). The speciality of FQ-PCR assay in testing clinical specimens was 95%.ConclusionThe FQ-PCR assay performed in a hermetially sealed tube is simple, rapid, anticontaminative. This assay is higher in sensitivity and speciality than smear and culture and can be used as an effective method for diagnosis of tuberculosis.