Rapid profiling of drug-resistant bacteria using DNA-binding dyes and a nanopore-based DNA sequencer

Spread of drug-resistant bacteria is a serious problem worldwide. We thus designed a new sequence-based protocol that can quickly identify bacterial compositions of clinical samples and their drug-resistance profiles simultaneously. Here we utilized propidium monoazide (PMA) that prohibits DNA ampli...

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Veröffentlicht in:Scientific reports 2021-02, Vol.11 (1), p.3436-3436, Article 3436
Hauptverfasser: Ohno, Ayumu, Umezawa, Kazuo, Asai, Satomi, Kryukov, Kirill, Nakagawa, So, Miyachi, Hayato, Imanishi, Tadashi
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Sprache:eng
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Zusammenfassung:Spread of drug-resistant bacteria is a serious problem worldwide. We thus designed a new sequence-based protocol that can quickly identify bacterial compositions of clinical samples and their drug-resistance profiles simultaneously. Here we utilized propidium monoazide (PMA) that prohibits DNA amplifications from dead bacteria, and subjected the original and antibiotics-treated samples to 16S rRNA metagenome sequencing. We tested our protocol on bacterial mixtures, and observed that sequencing reads derived from drug-resistant bacteria were significantly increased compared with those from drug-sensitive bacteria when samples were treated by antibiotics. Our protocol is scalable and will be useful for quickly profiling drug-resistant bacteria.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-021-82903-z