CRISPR/Cas12a-assisted rapid identification of key beer spoilage bacteria

Beer spoilage bacteria present severe challenges for the beer fermentation industry. Here, we establish a rapid, specific and highly portable CRISPR/Cas12a-based key beer spoilage bacterial scanning method, dubbed CRISPRBeer Scan. With the variable 16S rDNA segments of these major spoilage microbe species serving as targets, highly efficient and specific crRNAs were determined. By coupling with recombinase polymerase isothermal amplification, CRISPR-Beer Scan could detect as low as 10 copies of DNA targets. Indeed, trace amounts of target genome DNAs can be accurately identified within a mixture of genome DNAs conditioned in beer. Moreover, the extracted genome DNA samples can be conveniently distinguished through the visual fluorescent signals excited by blue light in 45 min with CRISPR-Beer Scan Method. Taken together, the CRISPR-Beer Scan represents a widely applicable microbe-monitoring platform for not only breweries, but also other industries.

Automated summary

A rapid, specific, and highly portable CRISPRBeer Scan method was established for detecting beer spoilage bacteria, achieving highly efficient and specific crRNAs targeting variable 16S rDNA segments. This method can detect as low as 10 copies of DNA targets and accurately identify trace amounts of target genome DNAs within a mixture of genome DNAs in beer. The extracted genome DNA samples can be conveniently distinguished through visual fluorescent signals excited by blue light in just 45 minutes, making CRISPR-Beer Scan a widely applicable microbe-monitoring platform for breweries and other industries.