Single-nucleotide polymorphism-based differentiation and drug resistance detection in Mycobacterium tuberculosis from isolates or directly from sputum
Single-nucleotide polymorphism-based differentiation and drug resistance detection in Mycobacterium tuberculosis from isolates or directly from sputum
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DOI：
10.1111/j.1469-0691.2004.01034.x
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发表时间：
2005-02-01
影响因子：
14.2
通讯作者：
Gharbia, S
中科院分区：
文献类型：
--
作者：
Arnold, C;Westland, L;Gharbia, S
应用焦磷酸测序技术对123份结核分枝杆菌DNA提取物和灭活痰标本进行了检测。在99份结核分枝杆菌单核苷酸多态性（SNP）研究样本中，68%的异烟肼耐药分离株在katG密码子315处存在AGC -> ACC突变，导致先前与异烟肼耐药相关的Ser -> Thr置换。在利福平耐药分离株中，92%在rpoB密码子516、531或526处显示SNP。灭活痰液样本和DNA提取物都可以通过焦磷酸测序进行分析，并且该方法能够快速区分密切相关的M.结核复合体（M.结核分枝杆菌、牛分枝杆菌、非洲分枝杆菌、卡内蒂分枝杆菌和田鼠分枝杆菌），除了M.结核分枝杆菌（M. canetti和两个M之一。非洲菌株。这种低成本、高通量的技术可用于快速筛选耐药性，并取代目前用于物种鉴定的一些耗时的测试。
The rapid technique of pyrosequencing was used to examine 123 samples (in the form of DNA extracts and inactivated sputum) of Mycobacterium spp. Of 99 Mycobacterium tuberculosis samples investigated for single-nucleotide polymorphisms (SNPs), 68% of isoniazid-resistant isolates analysed had an AGC --> ACC mutation in katG at codon 315, resulting in the Ser --> Thr substitution associated previously with isoniazid resistance. Of the rifampicin-resistant isolates, 92% showed SNPs in rpoB at codons 516, 531 or 526. Inactivated sputum samples and DNA extracts could both be analysed by pyrosequencing, and the method was able to differentiate rapidly between the closely related species of the M. tuberculosis complex (M. tuberculosis, Mycobacterium bovis, Mycobacterium africanum, Mycobacterium canetti and Mycobacterium microti), except between M. tuberculosis, M. canetti and one of two M. africanum strains. This low-cost, high-throughput technique could be used as a rapid screen for drug resistance and as a replacement for some of the time-consuming tests used currently for species identification.