DETECTION OF RIFAMPICIN-RESISTANCE MUTATIONS IN MYCOBACTERIUM-TUBERCULOSIS
DETECTION OF RIFAMPICIN-RESISTANCE MUTATIONS IN MYCOBACTERIUM-TUBERCULOSIS
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DOI：
10.1016/0140-6736(93)90417-f
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发表时间：
1993-03-13
期刊：
影响因子：
168.9
通讯作者：
BODMER, T
中科院分区：
文献类型：
--
作者：
TELENTI, A;IMBODEN, P;BODMER, T
结核病的控制受到结核分枝杆菌耐药性广泛出现的威胁。了解耐药性的分子基础可能会导致开发新的快速诊断耐药性的方法。我们着手确定利福平耐药的分子基础，利福平是用于治疗结核病的多药方案的主要组成部分。利福平耐药涉及RNA聚合酶的改变。克隆了编码RNA聚合酶β亚基（rpoB）的基因。从这个基因的序列信息被用来设计引物直接扩增和测序的411 bp的rpoB片段从122个结核分枝杆菌分离株。涉及8个保守氨基酸的突变被确定在64的66个不同地理来源的利福平耐药菌株，但在56个敏感菌株。所有突变都聚集在23个氨基酸的区域内。因此，由rpoB基因编码的有限数量的高度保守氨基酸的取代似乎是结核分枝杆菌对利福平的“单步”高水平耐药的分子机制。这些信息被用来开发一种策略（聚合酶链反应-单链构象多态性），允许有效地检测所有已知的利福平耐药突变体，这些发现提供了快速检测利福平耐药性的基础，耐多药结核病的标志。
Control of tuberculosis is threatened by widespread emergence of drug resistance in Mycobacterium tuberculosis. Understanding the molecular basis of resistance might lead to development of novel rapid methods for diagnosing drug resistance. We set out to determine the molecular basis of resistance to rifampicin, a major component of multidrug regimens used for treating tuberculosis.Resistance to rifampicin involves alterations of RNA polymerase. The gene that encodes the RNA polymerase subunit beta (rpoB) was cloned. Sequence information from this gene was used to design primers for direct amplification and sequencing of a 411 bp rpoB fragment from 122 isolates of M tuberculosis. Mutations involving 8 conserved aminoacids were identified in 64 of 66 rifampicin-resistant isolates of diverse geographical origin, but in none of 56 sensitive isolates. All mutations were clustered within a region of 23 aminoacids. Thus, substitution of a limited number of highly conserved aminoacids encoded by the rpoB gene appears to be the molecular mechanism responsible for ''single step'' high-level resistance to rifampicin in M tuberculosis. This information was used to develop a strategy (polymerase chain reaction-single-strand conformation polymorphism) that allowed efficient detection of all known rifampicin-resistant mutants.These findings provide the basis for rapid detection of rifampicin resistance, a marker of multidrug-resistant tuberculosis.