实时荧光定量PCR检测痰中结核分枝杆菌临床评价

The clinical evaluation of real-time FQ-PCR in the examination of Mycobacterium tuberculosis in sputum

目的评价实时荧光定量聚合酶链反应（FQ-PCR）检测临床痰标本中结核分枝杆菌诊断肺结核的意义。方法收集我院2011年9月-2013年8月期间收治的126例确诊为肺结核的患者和80例非结核性呼吸道疾病患者的痰液标本，分别采用漂浮集菌涂片镜检、BACT／ALERT3D快速培养法和FQ-PCR法检测痰中结核杆菌，分析检出结果。结果126例确诊为肺结核患者的痰标本中对结核分枝菌杆的检验以FQ-PCR检出率最高达到53．2％，高于快速培养法的46．8％（P〈0．05）和痰漂浮集菌涂片镜检法的32．5％（P〈0．01）。FQ-PCR与痰培养法总体一致性为95．1％，K=0．88。对痰涂片阳性标本，两者敏感性差异无统计学意义（P〉0．05）；对痰涂片阴性标本，FQ-PCR敏感性明显高于快速培养法（P〈0．01）。结论FQ-PCR具有快速、敏感、特异、可定量和自动化的优点，值得应用于痰中结核杆菌的检测及肺结核的诊断。

Objective To evaluate the clinical value of real-time fluorescence quantitative PCR （FQ-PCR） on the ex- amination of Mycobacterium tuberculosis in the sputum for diagnosis of pulmonary tuberculosis （FIB）. Methods 126 sputum samples from FIB patients and 80 sputum samples from non-pulmonary tuberculosis patients from Sep 2011 to Aug 2013 were selected from our hospital. All samples were detected by flotation bacteria smear microscopy, BACT/ALERT3D culture method and FQ -PCR for the examination of Mycobacterium tuberculosis in sputum. The results were analysed. Results For 126 con- firmed FIB samples, FQ-PCR showed the highest positive rate （53.2%）, higher than rapid culture method （46.8% ,P〈0.05） and flotation smear （32.5%,P〈0.01）. The FQ-PCR indicated 95.1% consisting with the culture assay. For the smear-positive specimens, the TB detection sensitivity of the FQ-PCR assay was similar with the BACT/ALERT3D culture method（P〉0.05）. For the smear-negative specimens, the TB detection sensitivity of the FQ-PCR assay was higher than the BACT/ALERT3D culture system （P〈0.01）. Conclusion FQ-PCR assay is a fast, sensitive, specific, quantitative and automatic method in the detec- tion of sputum Mycobacterium tuberculosis, with the value of clinical application in the diagnosis of FIB.