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bioRxiv · 10.1101/2020.10.09.332585

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

Ohno, A.·Umezawa, K.·Asai, S.·Kryukov, K.·Nakagawa, S.·Miyachi, H.·Imanishi, T.
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Abstract

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.

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Ohno, A., Umezawa, K., Asai, S., Kryukov, K., Nakagawa, S., Miyachi, H., Imanishi, T.. 2020-10-09. Rapid profiling of drug-resistant bacteria using DNA-binding dyes and a nanopore-based DNA sequencer. https://doi.org/10.1101/2020.10.09.332585

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