Identification of Mycobacterium abscessus species and subspecies using the Cas12a/sgRNA-based nucleic acid detection platform

Eur J Clin Microbiol Infect Dis. 2020 Mar;39(3):551-558. doi: 10.1007/s10096-019-03757-y. Epub 2019 Nov 28.

Abstract

The rapidly growing mycobacterium Mycobacterium abscessus is a clinically important organism causing pulmonary and skin diseases. The M. abscessus complex is comprised of three subspecies: M. abscessus subsp. abscessus, M. abscessus subsp. massiliense, and M. abscessus subsp. bolletii. Here, we aimed to develop a Cas12a/sgRNA-based nucleic acid detection platform to identify M. abscessus species and subspecies. By designing specific sgRNA probes targeting rpoB and erm(41), we demonstrated that M. abscessus could be differentiated from other major mycobacterial species and identified at the subspecies level. Using this platform, a total of 38 clinical M. abscessus isolates were identified, 18 as M. abscessus subsp. abscessus and 20 as M. abscessus subsp. massiliense. We concluded that the Cas12a/sgRNA-based nucleic acid detection platform provides an easy-to-use, quick, and cost-effective approach for identification of M. abscessus species and subspecies.

Keywords: FnCas12a; Identification; Mycobacterium abscessus subsp. abscessus; Mycobacterium abscessus subsp. bolletii; Mycobacterium abscessus subsp. massiliense; sgRNA probe.

MeSH terms

  • Bacterial Proteins / genetics*
  • CRISPR-Associated Proteins / genetics*
  • CRISPR-Cas Systems
  • DNA, Bacterial
  • Endodeoxyribonucleases / genetics*
  • Genes, Bacterial
  • Humans
  • Molecular Typing / methods*
  • Mycobacterium Infections, Nontuberculous / diagnosis*
  • Mycobacterium Infections, Nontuberculous / microbiology*
  • Mycobacterium abscessus / classification*
  • Mycobacterium abscessus / genetics*
  • Phylogeny
  • Polymerase Chain Reaction
  • RNA, Guide, CRISPR-Cas Systems
  • Sequence Analysis, DNA
  • Workflow

Substances

  • Bacterial Proteins
  • CRISPR-Associated Proteins
  • DNA, Bacterial
  • Cas12a protein
  • Endodeoxyribonucleases