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FLASH: a next-generation CRISPR diagnostic for multiplexed detection of antimicrobial resistance sequences.
- Author(s): Quan, Jenai
- Langelier, Charles
- Kuchta, Alison
- Batson, Joshua
- Teyssier, Noam
- Lyden, Amy
- Caldera, Saharai
- McGeever, Aaron
- Dimitrov, Boris
- King, Ryan
- Wilheim, Jordan
- Murphy, Maxwell
- Ares, Lara Pesce
- Travisano, Katherine A
- Sit, Rene
- Amato, Roberto
- Mumbengegwi, Davis R
- Smith, Jennifer L
- Bennett, Adam
- Gosling, Roly
- Mourani, Peter M
- Calfee, Carolyn S
- Neff, Norma F
- Chow, Eric D
- Kim, Peter S
- Greenhouse, Bryan
- DeRisi, Joseph L
- Crawford, Emily D
- et al.
Published Web Location
https://doi.org/10.1093/nar/gkz418Abstract
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.
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