Department of Microbiology, KGMU, Lucknow, Uttar Pradesh, India.
Int J Mycobacteriol. 2024 Jul 1;13(3):331-336. doi: 10.4103/ijmy.ijmy_139_24. Epub 2024 Sep 14.
Phenotypic drug susceptibility testing (DST) is considered the gold standard for detecting linezolid (LZD) resistance in Mycobacterium tuberculosis (MTB), but it is time-consuming. Nanopore sequencing offers a potentially faster alternative approach. This study evaluated the agreement between phenotypically detected LZD resistance and mutations in the rrl and rplC genes of MTB isolates using nanopore sequencing.
Consecutive drug-resistant MTB isolates from pulmonary samples collected in 2021 underwent liquid culture (LC) DST for LZD. All resistant isolates and an equal number of susceptible isolates were subjected to targeted sequencing of the rrl and rplC genes using nanopore technology.
Sequencing identified a C154R mutation in the rplC gene in only one LZD-resistant isolate. No mutations were detected in the rrl gene. The agreement between sequencing and LC-DST for detecting LZD resistance was poor (Cohen's kappa: 0.03571, 95% confidence interval [CI]: -0.034-0.105). Additionally, no significant association was found between LZD resistance and clinical or microbiological outcomes at 6-month follow-up.
This study revealed a considerable discrepancy between phenotypic and genotypic detection of LZD resistance in MTB. Further research is needed to better understand the genetic mechanisms underlying LZD resistance and to develop reliable molecular diagnostics for rapid resistance detection.
表型药敏试验(DST)被认为是检测结核分枝杆菌(MTB)利奈唑胺(LZD)耐药性的金标准,但它耗时较长。纳米孔测序提供了一种潜在的更快替代方法。本研究评估了纳米孔测序检测到的表型 LZD 耐药性与 MTB 分离株 rrl 和 rplC 基因中的突变之间的一致性。
对 2021 年采集的肺部耐药 MTB 分离株进行连续液体培养(LC)DST 检测 LZD。对所有耐药分离株和等量的敏感分离株进行 rrl 和 rplC 基因的靶向纳米孔测序。
测序仅在一个 LZD 耐药分离株中鉴定到 rplC 基因中的 C154R 突变。rrl 基因未检测到突变。测序与 LC-DST 检测 LZD 耐药性之间的一致性较差(Cohen's kappa:0.03571,95%置信区间 [CI]:-0.034-0.105)。此外,在 6 个月随访时,LZD 耐药性与临床或微生物学结局之间未发现显著相关性。
本研究揭示了 MTB 中表型和基因型检测 LZD 耐药性之间存在相当大的差异。需要进一步研究以更好地了解 LZD 耐药性的遗传机制,并开发用于快速耐药检测的可靠分子诊断。