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首页> 外文期刊>Journal of Clinical Microbiology >PCR-Based Rapid Identification System Using Bridged Nucleic Acids for Detection of Clarithromycin-Resistant Mycobacterium avium-M. intracellulare Complex Isolates
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PCR-Based Rapid Identification System Using Bridged Nucleic Acids for Detection of Clarithromycin-Resistant Mycobacterium avium-M. intracellulare Complex Isolates

机译：基于桥式核酸的基于PCR的快速鉴定系统，用于检测抗克拉霉素的鸟分枝杆菌M.细胞内复合物分离物

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The nontuberculous mycobacteria (NTM) cause miscellaneous disorders in humans, especially in the lungs, which present with a variety of radiological features. To date, knowledge of the pathogenic role of the Mycobacterium avium-intracellulare complex (MAC) in the human lung and the definitive criteria for initiating multidrug therapy are still lacking. However, there is little doubt that clarithromycin is the most efficacious drug among the various treatment regimens for lung NTM. In this study, with the use of a bridged nucleic acid (BNA) probe a detection system based on a real-time PCR (BNA-PCR) for the identification of the point mutations at position 2058 or 2059 in domain V of the 23S rRNA gene responsible for clarithromycin resistance was developed and has been assessed using MAC isolates from clinical samples. Out of 199 respiratory specimens, the drug susceptibility test demonstrated 12 strains resistant to clarithromycin, while the BNA-PCR showed 8 strains carrying the point mutation at position 2058 or 2059 of the 23S rRNA gene. This system revealed that there were mycobacterial strains resistant to clarithromycin which do not carry previously identified resistance genes. This paper documents a novel system for detecting clarithromycin-resistant strains and demonstrates that although these mutations are tacitly assumed to account for >90% of the reported resistant mutants, there is a significant fraction of resistant mutants that do not harbor these mutations. Therefore, unknown mechanisms affecting clarithromycin resistance remain to be elucidated.
机译：非结核分枝杆菌（NTM）会导致人类，尤其是肺部出现各种放射学特征的其他疾病。迄今为止，仍缺乏关于鸟分枝杆菌-细胞内复合物（MAC）在人肺中的致病作用的知识以及启动多药治疗的明确标准。但是，毫无疑问，克拉霉素是肺NTM各种治疗方案中最有效的药物。在这项研究中，使用桥接核酸（BNA）探针的检测系统基于实时PCR（BNA-PCR），用于鉴定23S rRNA域V中2058或2059处的点突变已经开发了负责克拉霉素抗性的基因，并已使用临床样品中的MAC分离物对其进行了评估。在199个呼吸道标本中，药物敏感性测试显示12株对克拉霉素具有抗性，而BNA-PCR显示8株在23S rRNA基因的2058或2059位置带有点突变。该系统显示存在对克拉霉素具有抗性的分枝杆菌菌株，其不携带先前鉴定的抗性基因。这篇论文记录了一种新型的检测克拉霉素抗性菌株的系统，并证明尽管默认假定这些突变占报道的抗性突变体的90％以上，但仍有相当一部分不包含这些突变的抗性突变体。因此，影响克拉霉素抗性的未知机制仍有待阐明。

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