检验医学 ›› 2014, Vol. 29 ›› Issue (10): 1037-1040.DOI: 10.3969/j.issn.1673-8640.2014.10.013

• 技术研究与评价·论著 • 上一篇    下一篇

分子杂交法快速鉴定结核分枝杆菌和非结核分枝杆菌的初步评价

于霞, 尚媛媛, 赵立平, 董玲玲, 马异峰, 王晓波, 黄海荣   

  1. 北京市结核病胸部肿瘤研究所 首都医科大学附属北京胸科医院, 北京 101149
  • 收稿日期:2013-05-30 出版日期:2014-10-30 发布日期:2014-10-22
  • 通讯作者: 黄海荣,联系电话:010-89509159。
  • 作者简介:于 霞,女,1983年生,硕士,研究实习员,主要从事结核分枝杆菌的实验室诊断和耐药研究。
  • 基金资助:

    北京市重大科技攻关项目(D121100003012001)

Primary evaluation of molecular hybridization method for the identification of Mycobacterium tuberculosis and non-tuberculosis Mycobacterium

YU Xia, SHANG Yuanyuan, ZHAO Liping, DONG Lingling, MA Yifeng, WANG Xiaobo, HUANG Hairong.   

  1. Beijing Tuberculosis and Thoracic Tumor Institute, Beijing Chest Hospital Affiliated to Capital Medical University, Beijing 101149,China
  • Received:2013-05-30 Online:2014-10-30 Published:2014-10-22

摘要:

目的 评价结核分枝杆菌鉴定试剂盒(分子杂交法)进行分枝杆菌菌种初筛的准确性,并探讨如何快速准确地鉴定结核分枝杆菌复合群(MTC)和非结核分枝杆菌(NTM)混合感染标本。方法 采用分子杂交法对200株结核分枝杆菌临床分离株进行菌种鉴定。人工制备结核分枝杆菌和胞内分枝杆菌、结核分枝杆菌和脓肿分枝杆菌、结核分枝杆菌和偶发分枝杆菌的混合感染模型,评价基因测序法、芯片法和Hain test在检测分枝杆菌混合感染中的准确性。结果 以对硝基苯甲酸(PNB)分离鉴定结果为参照,分子杂交法的敏感性和特异性分别为65.12%(65/86)和98.25%(112/114)。对于人工制备的13种不同比例混合的H37Rv和NTM,芯片法和Hain test均可以准确鉴定出混合感染标本中的混合菌种,而基因测序结果表明,H37Rv:偶发分枝杆菌的比例在10%~90%范围内才能区分出混合感染的类型。结论 分子杂交法对于结核分枝杆菌诊断有较高的特异性,一旦经该法鉴定为NTM,则具有很高的诊断意义。明确标本中存在混合感染后可以采用芯片法或Hain test对临床上高致病的分枝杆菌进行菌种水平的鉴定。

关键词: 结核分枝杆菌, 非结核分枝杆菌, 混合感染, 分子杂交法

Abstract:

Objective To evaluate the accuracy of Mycobacterial species identification kit (molecular hybridization method), and to investigate the rapid and accurate approaches for the identification of mixed infection caused by Mycobacterium tuberculosis complex (MTC) and non-tuberculosis Mycobacterium(NTM). Methods A total of 200 clinical Mycobacterium isolates were identified by molecular hybridization method. The mixed infection was simulated by the mixture of Mycobacterium tuberculosis and Mycobacterium intracellular, Mycobacterium tuberculosis and Mycobacterium kansasii, Mycobacterium tuberculosis and Mycobacterium fortuitum. The gene sequencing, gene chip array and Hain test were used to evaluate the accuracy of Mycobacterium-mixed infection. Resutls As reference to pnitrobenzoic acid(PNB) tests, the sensitivity and specificity of molecular hybridization method were 65.12%(65/86) and 98.25%(112/114), respectively. Among 13 different proportions of Mycobacterium tuberculosis and NTM, gene chip array and Hain test accurately detected all of the mixed infections. Gene sequencing can only detect the mixed infection caused by Mycobacterium tuberculosis and Mycobacterium fortuitum at a range from 10% to 90%. Conclusions The specificity of molecular hybridization method for the identification of Mycobacterium tuberculosis is high. The kit following the method for NTM has a significant diagnosis value. Once the mixed infection is identified in the culture, gene chip array and Hain test can identify the species of mixed culture for the common pathogenic Mycobacterium.

Key words: Non-tuberculosis , Mixed infection, Molecular hybridization method

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