Development of a rapid detection method for the macrolide resistance gene in Mycobacterium avium using the amplification refractory mutation system–loop-mediated isothermal amplification method

Takayuki Inagaki, Shoki Asahi, Kenji Ogawa, Taku Nakagawa, Teruko Ohkura, Yukari Osada, Toshiaki Nikai, Kiyofumi Yamada, Tetsuya Yagi, Kei Ichi Uchiya

研究成果: ジャーナルへの寄稿学術論文査読

2 被引用数 (Scopus)

抄録

Macrolide antibiotics such as clarithromycin (CLR) and azithromycin are the key drugs used in multidrug therapy for Mycobacterium avium complex (MAC) diseases. For these antibacterial drugs, drug susceptibility has been correlated with clinical response in MAC diseases. We have previously demonstrated the correlation between drug susceptibility and mutations in the 23S rRNA gene, which confers resistance to macrolides. Herein, we developed a rapid detection method using the amplification refractory mutation system (ARMS)–loop-mediated isothermal amplification (LAMP) technique to identify mutations in the 23S rRNA gene of M. avium. We examined the applicability of the ARMS–LAMP method to genomic DNA extracted from six genotypes of M. avium clinical isolates. The M. avium isolates were classified into 21 CLR-resistant and 9 CLR-susceptible strains based on the results of drug susceptibility tests; the 23S rRNA genes of these strains were sequenced and analyzed using the ARMS–LAMP method. Sequence analysis revealed that the 9 CLR-sensitive strains were wild-type strains, whereas the 21 CLR-resistant strains comprised 20 mutant-type strains and one wild-type strain. Using ARMS–LAMP, no amplification from genomic DNAs of the 10 wild-type strains was observed using the mutant-type mismatch primer sets (MTPSs); however, amplification from the 20 mutant-type strain DNAs was observed using the MTPSs. The rapid detection method developed by us integrates ARMS–LAMP with a real-time turbidimeter, which can help determine drug resistance in a few hours. In conclusion, ARMS–LAMP might be a new clinically beneficial technology for rapid detection of mutations.

本文言語英語
ジャーナルMicrobiology spectrum
12
4
DOI
出版ステータス出版済み - 04-2024
外部発表はい

All Science Journal Classification (ASJC) codes

  • 生理学
  • 生態学
  • 免疫学および微生物学一般
  • 遺伝学
  • 微生物学(医療)
  • 細胞生物学
  • 感染症

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