Detection of rpoB, katG and inhA gene mutations in Mycobacterium tuberculosis clinical isolates from Chongqing as determined by microarray
Detection of rpoB, katG and inhA gene mutations in Mycobacterium tuberculosis clinical isolates from Chongqing as determined by microarray
复制标题
微阵列检测重庆结核分枝杆菌临床分离株rpoB、katG和inhA基因突变
DOI：
10.1111/j.1469-0691.2010.03267.x
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发表时间：
2010-11-01
影响因子：
14.2
通讯作者：
Fu, W.
中科院分区：
文献类型：
--
作者：
Yao, C.;Zhu, T.;Fu, W.
耐多药结核分枝杆菌的出现对结核病控制规划的威胁越来越大。采用传统方法进行结核分枝杆菌复合群分离株的药敏试验至少需要14天。如果使用分子分析，这可以显著降低。DNA测序是检测突变的好方法，但由于其相对较高的成本而不能常规使用。设计了一种敏感、特异的基因芯片，用于检测利福平耐药决定区rpoB和与异烟肼（INH）耐药相关的katG和inhA位点的突变。使用一组含有13种不同rpoB基因型、katG密码子315内的两种突变基因型和inhA处的一种突变基因型的结核分枝杆菌分离株来验证微阵列。结果表明，100%的利福平耐药M。重庆地区分离的结核菌株存在rpoB突变，531-Ser和526-His是最常见的位置置换。在总共50株INH耐药分离株中，82%具有katG315突变，18%具有inhA突变。所有通过微阵列方法检测到的突变也通过常规DNA测序证实。结果表明，基因芯片技术是一种高效、特异的检测方法，可用于利福平和异烟肼耐药的快速检测。
P>The emergence of multidrug-resistance Mycobacterium tuberculosis is an increasing threat to tuberculosis control programmes. Susceptibility testing of Mycobacterium tuberculosis complex isolates by traditional methods requires a minimum of 14 days. This can be reduced significantly if molecular analysis is used. DNA sequencing is a good method for detecting mutation, but cannot be used routinely because of its relatively high cost. A sensitive and specific microarray has been designed to detect mutations in the rifampin resistance determining region of rpoB and loci in katG and inhA associated with isoniazid (INH) resistance. A panel of Mycobacterium tuberculosis isolates containing 13 different rpoB genotypes, two mutation genotypes within codon 315 of katG and one mutation genotypes at inhA was used to validate the microarray. The results obtained indicate that 100% of rifampicin-resistant M. tuberculosis strains isolated in Chongqing had rpoB mutations, with 531-Ser and 526-His being the most common positions substituted. Of the total 50 INH resistant isolates, 82% had a katG315 mutation and 18% had an inhA mutation. All the mutations detected by the microarray method were also confirmed by conventional DNA sequencing. It is demonstrated that the microarray is an efficient, specialized technique and can be used as a rapid method for detecting rifampin and isoniazid resistance.