Molecular Diagnosis of Fluoroquinolone Resistance in Mycobacterium tuberculosis
Molecular Diagnosis of Fluoroquinolone Resistance in Mycobacterium tuberculosis
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DOI：
10.1128/aac.04058-14
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发表时间：
2015-03-01
影响因子：
4.9
通讯作者：
Aubry, Alexandra
中科院分区：
文献类型：
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作者：
Bernard, Christine;Veziris, Nicolas;Aubry, Alexandra
由于使用氟喹诺酮类药物（FQ），出现了对FQ的耐药性，导致几乎无法治疗和广泛耐药的结核病病例。DNA促旋酶的突变代表了FQ抗性的主要机制。全面了解耐药（FQ（r））临床分离株中发现的突变模式及其比例，对于改进检测结核分枝杆菌FQ耐药的分子方法至关重要。在这项研究中，我们回顾了2007年至2012年法国国家分枝杆菌参考中心分离株中FQ耐药性的检测，目的是评估PCR测序在现实生活中的性能。gyrA和gyrB测序，对M.用参比比例法比较结核临床分离株对2 mg/L氧氟沙星的敏感性。共分析了605株分离株，其中50%为多药耐药株。在研究期间，多药耐药（MDR）菌株中FQ（r）菌株的增加令人担忧（8%至30%）。大多数（78%）gyrA突变的分离株是FQ（r），而只有36%的gyrB突变的分离株是FQ（r）。只有12%的FQ（r）分离株在gyrB中有单一突变。组合的gyrA和gyrB测序导致>93%的检测耐药性的灵敏度。通过对gyrA和gyrB测序的4例假阳性和5例假阴性结果的分析，说明了参考比例法的实际局限性。我们的数据强调需要联合gyrA和gyrB测序的调查FQ敏感性在M。结核病和挑战的有效性，目前的表型为基础的方法作为诊断金标准，用于确定FQ耐药。
As a consequence of the use of fluoroquinolones (FQ), resistance to FQ has emerged, leading to cases of nearly untreatable and extensively drug-resistant tuberculosis. Mutations in DNA gyrase represent the main mechanism of FQ resistance. A full understanding of the pattern of mutations found in FQ-resistant (FQ(r)) clinical isolates, and of their proportions, is crucial for improving molecular methods for the detection of FQ resistance in Mycobacterium tuberculosis. In this study, we reviewed the detection of FQ resistance in isolates addressed to the French National Reference Center for Mycobacteria from 2007 to 2012, with the aim of evaluating the performance of PCR sequencing in a real-life context. gyrA and gyrB sequencing, performed prospectively on M. tuberculosis clinical isolates, was compared for FQ susceptibility to 2 mg/liter ofloxacin by the reference proportion method. A total of 605 isolates, of which 50% were multidrug resistant, were analyzed. The increase in FQ(r) strains among multidrug-resistant (MDR) strains during the time of the study was alarming (8% to 30%). The majority (78%) of the isolates with gyrA mutations were FQ(r), whereas only 36% of those with gyrB mutations were FQ(r). Only 12% of the FQ(r) isolates had a single mutation in gyrB. Combined gyrA and gyrB sequencing led to >93% sensitivity for detecting resistance. The analysis of the four false-positive and the five false-negative results of gyrA and gyrB sequencing illustrated the actual limitations of the reference proportion method. Our data emphasize the need for combined gyrA and gyrB sequencing in the investigation of FQ susceptibility in M. tuberculosis and challenge the validity of the current phenotype-based approach as the diagnostic gold standard for determining FQ resistance.