Detection of multidrug-resistance in Mycobacterium tuberculosis by phenotype- and molecular-based assays.

IF 4.6 2区 医学 Q1 MICROBIOLOGY
Laima Vasiliauskaitė, Zofia Bakuła, Edita Vasiliauskienė, Daiva Bakonytė, Przemysław Decewicz, Mikołaj Dziurzyński, Małgorzata Proboszcz, Edita Valerija Davidavičienė, Birutė Nakčerienė, Rafał Krenke, Tomas Kačergius, Petras Stakėnas, Tomasz Jagielski
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引用次数: 0

Abstract

Background: The whole-genome sequencing (WGS) is becoming an increasingly effective tool for rapid and accurate detection of drug resistance in Mycobacterium tuberculosis complex (MTBC). This approach, however, has still been poorly evaluated on strains from Central and Eastern European countries. The purpose of this study was to assess the performance of WGS against conventional drug susceptibility testing (DST) for the detection of multi-drug resistant (MDR) phenotypes among MTBC clinical strains from Poland and Lithuania.

Methods: The study included 208 MTBC strains (130 MDR; 78 drug susceptible), recovered from as many tuberculosis patients in Lithuania and Poland between 2018 and 2021. Resistance to rifampicin (RIF) and isoniazid (INH) was assessed by Critical Concentration (CC) and Minimum Inhibitory Concentration (MIC) DST as well as molecular-based techniques, including line-probe assay (LPA) and WGS. The analysis of WGS results was performed using bioinformatic pipeline- and software-based tools.

Results: The results obtained with the CC DST were more congruent with those by LPA compared to pipeline-based WGS. Software-based tools showed excellent concordance with pipeline-based analysis in prediction of RIF/INH resistance. The RIF-resistant strains demonstrated a relatively homogenous MIC distribution with the mode at the highest tested MIC value. The most frequent RIF-resistance conferring mutation was rpoB S450L. The mode MIC for INH was two-fold higher among double katG and inhA mutants than among single katG mutants. The overall rate of discordant results between all methods was calculated at 5.3%. Three strains had discordant results by both genotypic methods (LPA and pipeline-based WGS), one strain by LPA only, three strains by MIC DST, two strains by both MIC DST and pipeline-based WGS, and the remaining two strains showed discordant results with all three methods, compared to CC DST.

Conclusions: Considering MIC DST results, current CCs of the first-line anti-TB drugs might be inappropriately high and may need to be revised. Both molecular methods demonstrated 100% specificity, while pipeline-based WGS had slightly lower sensitivity for RIF and INH than LPA, compared to CC DST.

通过表型和分子测定法检测结核分枝杆菌的多重耐药性。
背景:全基因组测序(WGS)正日益成为快速准确检测结核分枝杆菌(MTBC)耐药性的有效工具。然而,这种方法在中欧和东欧国家的菌株中的评估结果还很不理想。本研究旨在评估 WGS 与传统药敏试验(DST)在检测波兰和立陶宛 MTBC 临床菌株耐多药(MDR)表型方面的性能:研究纳入了 208 株 MTBC 菌株(130 株 MDR;78 株药敏),这些菌株于 2018 年至 2021 年间从立陶宛和波兰的多位结核病患者中回收。对利福平(RIF)和异烟肼(INH)的耐药性通过临界浓度(CC)和最低抑制浓度(MIC)DST以及基于分子的技术(包括线探针测定(LPA)和WGS)进行评估。对 WGS 结果的分析是利用生物信息管道和软件工具进行的:结果:与基于管道的 WGS 相比,CC DST 得出的结果与 LPA 得出的结果更加一致。在预测 RIF/INH 耐药性方面,基于软件的工具与基于管道的分析具有极好的一致性。RIF 耐药菌株的 MIC 分布相对均匀,以最高测试 MIC 值为模式。最常见的 RIF 耐药性突变是 rpoB S450L。在双 katG 和 inhA 突变体中,INH 的 MIC 模式是单 katG 突变体的两倍。据计算,所有方法的总体结果不一致率为 5.3%。与 CC DST 相比,3 株菌株的两种基因分型方法(LPA 和基于管道的 WGS)结果不一致,1 株菌株仅使用 LPA,3 株菌株使用 MIC DST,2 株菌株使用 MIC DST 和基于管道的 WGS,其余 2 株菌株的三种方法结果均不一致:结论:考虑到 MIC DST 的结果,目前一线抗结核药物的 CC 值可能过高,需要进行修订。两种分子方法的特异性均为 100%,而与 CC DST 相比,基于管道的 WGS 对 RIF 和 INH 的敏感性略低于 LPA。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
8.60
自引率
0.00%
发文量
49
审稿时长
>12 weeks
期刊介绍: Annals of Clinical Microbiology and Antimicrobials considers good quality, novel and international research of more than regional relevance. Research must include epidemiological and/or clinical information about isolates, and the journal covers the clinical microbiology of bacteria, viruses and fungi, as well as antimicrobial treatment of infectious diseases. Annals of Clinical Microbiology and Antimicrobials is an open access, peer-reviewed journal focusing on information concerning clinical microbiology, infectious diseases and antimicrobials. The management of infectious disease is dependent on correct diagnosis and appropriate antimicrobial treatment, and with this in mind, the journal aims to improve the communication between laboratory and clinical science in the field of clinical microbiology and antimicrobial treatment. Furthermore, the journal has no restrictions on space or access; this ensures that the journal can reach the widest possible audience.
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