Genetic Determinants of Drug Resistance in Mycobacterium tuberculosis and Their Diagnostic Value
Genetic Determinants of Drug Resistance in Mycobacterium tuberculosis and Their Diagnostic Value
复制标题
DOI：
10.1164/rccm.201510-2091oc
复制
发表时间：
2016-09-01
影响因子：
24.7
通讯作者：
Murray, Megan
中科院分区：
文献类型：
--
作者：
Farhat, Maha R.;Sultana, Razvan;Murray, Megan
基本原理：分子诊断学的发展，检测结核分枝杆菌在临床样本中的存在和耐药性赋予突变，有望彻底改变病人的护理和中断通过确保早期诊断的传播。然而，这些工具需要鉴定对所有抗结核药物耐药的遗传决定因素。目的：为了确定所需的最佳分子方法，我们试图创建一个全面的耐药突变目录，并评估其诊断耐药的敏感性和特异性。方法：我们开发并验证了分子倒置探针，用于M中28个耐药基因座的DNA捕获和深度测序。结核我们使用这些探针对地理上不同的1，397例临床M.已知耐药表型的结核病分离株。我们确定了一组最小的突变来预测对一线和二线抗结核药物的耐药性，并在一个独立的数据集中验证了我们的预测。我们构建并试点了一个基于网络的数据库，提供公众访问的序列数据和predidtion tool. Measures和主要结果：预测的耐药性利福平和异烟肼超过90%的敏感性和特异性，但其他药物较低。诊断耐药所需的突变数量很大，对于研究的13种药物，在18个遗传位点上有238个突变。结核病耐药性诊断将需要考虑到高维度的突变检测。他们还支持这样的假设，即目前未知的遗传决定因素，可能通过全基因组测序，编码耐二线结核病药物。
Rationale: The development of molecular diagnostics that detect both the presence of Mycobacterium tuberculosis in clinical samples and drug resistance-conferring mutations promises to revolutionize patient care and interrupt transmission by ensuring early diagnosis. However, these tools require the identification of genetic determinants of resistance to the full range of antituberculosis drugs.Objectives: To determine the optimal molecular approach needed, we sought to create a comprehensive catalog of resistance mutations and assess their sensitivity and specificity in diagnosing drug resistance.Methods: We developed and validated molecular inversion probes for DNA capture and deep sequencing of 28 drug-resistance loci in M. tuberculosis. We used the probes for targeted sequencing of a geographically diverse set of 1,397 clinical M. tuberculosis isolates with known drug resistance phenotypes. We identified a minimal set of mutations to predict resistance to first- and second-line antituberculosis drugs and validated our predictions in an independent dataset. We constructed and piloted a web-based database that provides public access to the sequence data and predidtion tool.Measurements and Main Results: The predicted resistance to rifampicin and isoniazid exceeded 90% sensitivity and specificity but was lower for other drugs. The number of mutations needed to diagnose resistance is large, and for the 13 drugs studied it was 238 across 18 genetic loci.Conclusions: These data suggest that a comprehensive M. tuberculosis drug resistance diagnostic will need to allow for a high dimension of mutation detection. They also support the hypothesis that currently unknown genetic determinants, potentially discoverable by whole-genome sequencing, encode resistance to second-line tuberculosis drugs.