Evaluation of Pyrosequencing for Detecting Extensively Drug-Resistant Mycobacterium tuberculosis among Clinical Isolates from Four HighBurden Countries
Evaluation of Pyrosequencing for Detecting Extensively Drug-Resistant Mycobacterium tuberculosis among Clinical Isolates from Four HighBurden Countries
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DOI：
10.1128/aac.03614-14
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发表时间：
2015-01-01
影响因子：
4.9
通讯作者：
Rodwell, Timothy C.
中科院分区：
文献类型：
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作者：
Ajbani, Kanchan;Lin, Shou-Yean Grace;Rodwell, Timothy C.
可靠的分子诊断，检测与耐药性相关的特定突变，是快速鉴定和监测结核分枝杆菌菌株耐药性的有前途的技术。焦磷酸测序（PSQ）能够检测与一线和二线抗结核（TB）药物相关的突变，具有快速适应新突变鉴定的额外优势。该项目的目的是评估PSQ在预测来自印度、南非、摩尔多瓦和菲律宾的多药耐药和广泛耐药结核病（M/XDR-TB）临床分离株的表型耐药性方面的性能。通过PSQ试验运行了总计187个存档分离株，以鉴定M。结核病（通过IS 6110标记），并检测与M/XDR-TB相关的突变，在选定的基因座的小片段核苷酸。分子靶点包括katG、inhA启动子和异烟肼（INH）耐药的ahpC-oxyR基因间区;利福平（RIF）耐药的rpoB核心区;氟喹诺酮（FQ）耐药的gyrA;阿米卡星（AMK）、卷曲霉素（CAP）和卡那霉素（KAN）耐药的rrs。将PSQ数据与表型分枝杆菌生长指示管（MGIT）960药敏试验结果进行比较，以进行性能分析。PSQ法对INH（94%）、RIF（96%）、FQ（93%）、AMK（84%）、CAP（88%）和KAN（68%）的耐药性检测显示出良好的敏感性。该方法对INH的特异性为96%，对RIF、FQ、AMK和KAN的特异性为100%，对CAP的特异性为97%。PSQ是一种高效的诊断工具，可以揭示与一线和二线抗结核药物耐药性相关的特定核苷酸变化。这种方法有可能被链接到突变特异性的临床解释算法，快速治疗决策。
Reliable molecular diagnostics, which detect specific mutations associated with drug resistance, are promising technologies for the rapid identification and monitoring of drug resistance in Mycobacterium tuberculosis isolates. Pyrosequencing (PSQ) has the ability to detect mutations associated with first-and second-line anti-tuberculosis (TB) drugs, with the additional advantage of being rapidly adaptable for the identification of new mutations. The aim of this project was to evaluate the performance of PSQ in predicting phenotypic drug resistance in multidrug-and extensively drug-resistant tuberculosis (M/XDR-TB) clinical isolates from India, South Africa, Moldova, and the Philippines. A total of 187 archived isolates were run through a PSQ assay in order to identify M. tuberculosis (via the IS6110 marker), and to detect mutations associated with M/XDR-TB within small stretches of nucleotides in selected loci. The molecular targets included katG, the inhA promoter and the ahpC-oxyR intergenic region for isoniazid (INH) resistance; the rpoB core region for rifampin (RIF) resistance; gyrA for fluoroquinolone (FQ) resistance; and rrs for amikacin (AMK), capreomycin (CAP), and kanamycin (KAN) resistance. PSQ data were compared to phenotypic mycobacterial growth indicator tube (MGIT) 960 drug susceptibility testing results for performance analysis. The PSQ assay illustrated good sensitivity for the detection of resistance to INH (94%), RIF (96%), FQ (93%), AMK (84%), CAP (88%), and KAN (68%). The specificities of the assay were 96% for INH, 100% for RIF, FQ, AMK, and KAN, and 97% for CAP. PSQ is a highly efficient diagnostic tool that reveals specific nucleotide changes associated with resistance to the first- and second-line anti-TB drug medications. This methodology has the potential to be linked to mutation-specific clinical interpretation algorithms for rapid treatment decisions.