A novel DNA chromatography method to discriminate Mycobacterium abscessus subspecies and macrolide susceptibility

Mitsunori Yoshida, Sotaro Sano, Jung Yien Chien, Hanako Fukano, Masato Suzuki, Takanori Asakura, Kozo Morimoto, Yoshiro Murase, Shigehiko Miyamoto, Atsuyuki Kurashima, Naoki Hasegawa, Po Ren Hsueh, Satoshi Mitarai, Manabu Ato, Yoshihiko Hoshino

Research output: Contribution to journalArticlepeer-review

Abstract

Background: 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. Methods: 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. Finding: 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%. Interpretation: We developed a novel, rapid and simple DNA chromatography method for identification of MABC macrolide susceptibility with high accuracy. Funding: AMED, JSPS KAKENHI

Original languageEnglish
Article number103187
JournalEBioMedicine
Volume64
DOIs
Publication statusPublished - 2021 Feb

Keywords

  • ATS/ERS/ESCMID/IDSA guideline
  • DNA chromatography
  • Drug susceptibility
  • Non-tuberculosis mycobacteria
  • Subspecies discrimination
  • Whole genome sequencing

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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