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中国防痨杂志 ›› 2011, Vol. 33 ›› Issue (5): 263-266.

• 论著 • 上一篇    下一篇

结核/非结核分枝杆菌核酸快速检测方法临床应用价值

康丽军1 洪峰2 冯建兵3 刘威1 吴华1 易俊莉2 王甦民2   

  1. 1.北京老年医院; 2.北京结核病控制研究所; 3.北京洛奇临床检验所;
  • 收稿日期:2011-03-05 出版日期:2011-05-20 发布日期:2012-03-15
  • 通信作者: 洪峰 E-mail:hongfeng@bjhb.gov.cn

 Clinical value of real-time FQ-PCR assay for detection of Mycobacterium tuberculosis/non-tuberculous mycobacteria

Kang Lijun1, Hong Feng2, Feng Jianbing3, Liu Wei1, Wu Hua1, Yi Junli1, Yi Junli2, Wang Sumin2   

  1. 1.Beijing Old Hospital; Beijing 100095; China; 2.Beijing Research Institute for Tuberculosis Control; Beijing 100035; 3.Beijing Lawke HeaHh laboratory; Beijing 102206
  • Received:2011-03-05 Online:2011-05-20 Published:2012-03-15
  • Contact: Hong Feng E-mail:hongfeng@bjhb.gov.cn

摘要: 目的 评价结核/非结核分枝杆菌核酸快速检测方法的临床应用价值。 方法 应用实时荧光PCR技术对420例标本进行临床试验研究,并与抗酸杆菌涂片和分枝杆菌培养方法对照比较。 结果 结核分枝杆菌核酸检测灵敏度为48.5%(其中菌阳标本灵敏度为95.8%,菌阴标本灵敏度为14.5%),特异度99.3%,阳性预测值为99.1%,阴性预测值为55.5%;临床试验388例痰标本中检测出1例非结核分枝杆菌核酸阳性,经测序确认,准确率100%;同时对32例非结核分枝杆菌临床分离株进行核酸检测,并经测序确认,准确率100%。 结论 应用实时荧光PCR技术,能实现在1支管内同时进行结核分枝杆菌/非结核分枝杆菌的核酸检测。该方法具有简单快速、特异性好污染性少的特点。可作为实验室辅助检测结核/非结核分枝杆菌核酸方法之一。

关键词: 分枝杆菌, 结核, 分枝杆菌属, 核酸类, 聚合酶链反应

Abstract: Objective To evaluate the clinical diagnostic value of real-time FQ-PCR assay for the detection of Mycobacterium tuberculosis(MTB)/non-tuberculous mycobacteria (NTM). Methods 420 specimens were detected by real-time FQ-PCR, and the results were compared with those of AFB smear and culture.  Results The sensitivity of FQ-PCR was 48.5%, in which the sensitivity in bacterium-positive and bacterium-negative specimens were 95.8% and 14.5%, respectively. The specificity, positive predictive value and negative predictive value of FQ-PCR were 99.3%, 99.1% and 55.5%, respectively. 1 of 388 sputum specimens was NTM-positive by FQ-PCR, which was confirmed by DNA sequencing. 32 NTM isolates were confirmed by FQ-PCR and DNA sequencing. Their accuracy was 100%.  Conclusions The real-time FQ-PCR was a simple, rapid and specific assay for the detection ofMycobacterium tuberculosis /non-tuberculous mycobacteria in a sealed tube.

Key words: Mycobacterium tuberculosis, mycobacterum, nucleic acids, polymerase chain reaction