Department of Mycobacteriology, Leprosy Research Center, National Institute of Infectious Diseases, Toyko, Japan.
Medical Solutions Vehicle, Kaneka Co. Ltd., Hyogo, Japan.
EBioMedicine. 2021 Feb;64:103187. doi: 10.1016/j.ebiom.2020.103187. Epub 2021 Jan 11.
The clinical impact of infection with Mycobacterium (M.) abscessus complex (MABC), a group of emerging non-tuberculosis mycobacteria (NTM), is increasing. M. abscessus subsp. abscessus/bolletii frequently shows natural resistance to macrolide antibiotics, whereas M. abscessus subsp. massiliense is generally susceptible. Therefore, rapid and accurate discrimination of macrolide-susceptible MABC subgroups is required for effective clinical decisions about macrolide treatments for MABC infection. We aimed to develop a simple and rapid diagnostic that can identify MABC isolates showing macrolide susceptibility.
Whole genome sequencing (WGS) was performed for 148 clinical or environmental MABC isolates from Japan to identify genetic markers that can discriminate three MABC subspecies and the macrolide-susceptible erm(41) T28C sequevar. Using the identified genetic markers, we established PCR based- or DNA chromatography-based assays. Validation testing was performed using MABC isolates from Taiwan.
We identified unique sequence regions that could be used to differentiate the three subspecies. Our WGS-based phylogenetic analysis indicated that M. abscessus carrying the macrolide-susceptible erm(41) T28C sequevar were tightly clustered, and identified 11 genes that were significantly associated with the lineage for use as genetic markers. To detect these genetic markers and the erm(41) locus, we developed a DNA chromatography method that identified three subspecies, the erm(41) T28C sequevar and intact erm(41) for MABC in a single assay within one hour. The agreement rate between the DNA chromatography-based and WGS-based identification was 99·7%.
We developed a novel, rapid and simple DNA chromatography method for identification of MABC macrolide susceptibility with high accuracy.
AMED, JSPS KAKENHI.
分枝杆菌（M.）脓肿复合体（MABC）是一组新兴的非结核分枝杆菌（NTM），其感染的临床影响正在增加。M. abscessus 亚种。脓肿/bolletii 经常表现出对大环内酯类抗生素的天然耐药性，而 M. abscessus 亚种。massiliense 通常是敏感的。因此，需要快速准确地区分大环内酯类敏感的 MABC 亚群，以便为 MABC 感染的大环内酯类治疗做出有效的临床决策。我们旨在开发一种简单快速的诊断方法，可以识别显示大环内酯类敏感性的 MABC 分离株。
对来自日本的 148 株临床或环境 MABC 分离株进行全基因组测序（WGS），以鉴定可区分三种 MABC 亚种和大环内酯类敏感 erm(41)T28C 序列变异的遗传标记。使用鉴定的遗传标记，我们建立了基于 PCR 或 DNA 色谱的检测方法。使用来自台湾的 MABC 分离株进行验证测试。
我们确定了可用于区分三个亚种的独特序列区域。我们的 WGS 基于系统发育分析表明，携带大环内酯类敏感 erm(41)T28C 序列变异的 M. abscessus 紧密聚集，并确定了 11 个与谱系显著相关的基因，可作为遗传标记。为了检测这些遗传标记和 erm(41)基因座，我们开发了一种 DNA 色谱法，可在一个小时内的单次检测中鉴定 MABC 的三个亚种、erm(41)T28C 序列变异和完整的 erm(41)。DNA 色谱法与 WGS 鉴定的一致性率为 99.7%。
我们开发了一种新的、快速且简单的 DNA 色谱法，用于鉴定 MABC 对大环内酯类的敏感性，具有很高的准确性。
AMED、JSPS KAKENHI。
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