Médecins Sans Frontières, Mumbai, India.
Grant Medical College, Sir Jamshedjee Jeejebhoy Group of Hospitals, Mumbai, India.
Int J Tuberc Lung Dis. 2023 Jan 1;27(1):41-48. doi: 10.5588/ijtld.22.0138.
In high TB burden countries, access to drug susceptibility testing is a major bottleneck. Targeted next-generation sequencing (tNGS) is a promising technology for rapid resistance detection. This study assessed the role of tNGS for the diagnosis of drug-resistant TB (DR-TB). A total of 161 samples from bacteriologically confirmed TB cases were subjected to tNGS using the Deeplex Myc-TB kit and sequenced using the MiSeq platform. These samples were also processed for conventional phenotypic DST (pDST) using 13 drugs on Mycobacteria Growth Indicator Tube and line-probe assays (MTBDR and MTBDR). There were 146 DR-TB and 15 drug-susceptible TB (DS-TB) samples. About 70% of patients with DR-TB had no previous TB treatment history. Overall, 88.2% had rifampicin-resistant/multidrug-resistant TB (RR/MDR-TB), 58.5% pre-extensively drug-resistant TB (pre-XDR-TB) and 9.2% had XDR-TB as defined by the WHO (2020). Around 8% ( = 13) of samples were non-culturable; however, identified 8 were resistant to first and second-line drugs using tNGS. Resistance frequency was similar across methods, with discordance in drugs less reliable using pDST or with limited mutational representation within databases. Sensitivities were aligned with literature reports for most drugs. We observed 10% heteroresistance, while 75% of strains were of Lineages 2 and 3. Programme data supported tNGS in the diagnosis of DR-TB for early treatment using individualised regimens.
在结核病高负担国家，获得药敏检测是一个主要瓶颈。靶向下一代测序（tNGS）是一种快速耐药检测的有前途的技术。本研究评估了 tNGS 在耐药结核病（DR-TB）诊断中的作用。共对 161 例经细菌学证实的结核病病例的样本进行了 tNGS 检测，使用 Deeplex Myc-TB 试剂盒进行检测，并使用 MiSeq 平台进行测序。这些样本还使用 13 种药物在分枝杆菌生长指示管和线探针分析（MTBDR 和 MTBDR）上进行了常规表型药敏检测（pDST）。共有 146 例 DR-TB 和 15 例药物敏感结核病（DS-TB）样本。约 70%的 DR-TB 患者没有既往结核病治疗史。总体而言，88.2%的患者患有利福平耐药/耐多药结核病（RR/MDR-TB），58.5%的患者患有预广泛耐药结核病（pre-XDR-TB），9.2%的患者患有世界卫生组织（2020 年）定义的 XDR-TB。约 8%（=13）的样本不可培养；然而，使用 tNGS 鉴定出 8 种对一线和二线药物耐药。耐药频率与各种方法相似，pDST 中药物的不一致性或数据库中有限的突变代表不太可靠。对于大多数药物，敏感性与文献报道一致。我们观察到 10%的异质性耐药，而 75%的菌株属于谱系 2 和 3。项目数据支持使用个体化方案进行 DR-TB 的早期治疗中 tNGS 对 DR-TB 的诊断。
