FLASH: a next-generation CRISPR diagnostic for multiplexed detection of antimicrobial resistance sequences

J Quan, C Langelier, A Kuchta, J Batson… - Nucleic acids …, 2019 - academic.oup.com
J Quan, C Langelier, A Kuchta, J Batson, N Teyssier, A Lyden, S Caldera, A McGeever
Nucleic acids research, 2019academic.oup.com
The growing prevalence of deadly microbes with resistance to previously life-saving drug
therapies is a dire threat to human health. Detection of low abundance pathogen sequences
remains a challenge for metagenomic Next Generation Sequencing (NGS). We introduce
FLASH (Finding Low Abundance Sequences by Hybridization), a next-generation
CRISPR/Cas9 diagnostic method that takes advantage of the efficiency, specificity and
flexibility of Cas9 to enrich for a programmed set of sequences. FLASH-NGS achieves up to …
Abstract
The growing prevalence of deadly microbes with resistance to previously life-saving drug therapies is a dire threat to human health. Detection of low abundance pathogen sequences remains a challenge for metagenomic Next Generation Sequencing (NGS). We introduce FLASH (Finding Low Abundance Sequences by Hybridization), a next-generation CRISPR/Cas9 diagnostic method that takes advantage of the efficiency, specificity and flexibility of Cas9 to enrich for a programmed set of sequences. FLASH-NGS achieves up to 5 orders of magnitude of enrichment and sub-attomolar gene detection with minimal background. We provide an open-source software tool (FLASHit) for guide RNA design. Here we applied it to detection of antimicrobial resistance genes in respiratory fluid and dried blood spots, but FLASH-NGS is applicable to all areas that rely on multiplex PCR.
Oxford University Press