CRISPR/Cas12a-assisted rapid identification of key beer spoilage bacteria
CRISPR/Cas12a-assisted rapid identification of key beer spoilage bacteria
复制标题
CRISPR/Cas12a辅助快速鉴定啤酒关键腐败菌
DOI：
10.1016/j.ifset.2021.102854
复制
发表时间：
2021-10-24
影响因子：
6.6
通讯作者：
Leng, Qibin
中科院分区：
文献类型：
--
作者：
Meng, Qingzhou;Yang, Hongmei;Leng, Qibin
啤酒腐败菌对啤酒发酵工业提出了严峻的挑战。在这里，我们建立了一种快速，特异性和高度便携的基于CRISPR/Cas 12 a的关键啤酒腐败细菌扫描方法，称为CRISPRBeer Scan。以这些主要腐败微生物物种的可变16 S rDNA片段为靶点，确定了高效且特异的crRNA。通过与重组酶聚合酶等温扩增相结合，CRISPR-Beer Scan可以检测低至10个拷贝的DNA靶标。事实上，在啤酒中调节的基因组DNA的混合物中可以准确地鉴定痕量的靶基因组DNA。此外，采用CRISPR-Beer扫描方法，通过蓝光激发45 min的可见荧光信号，可以方便地对提取的基因组DNA样品进行区分。综上所述，CRISPR-Beer Scan代表了一个广泛适用的微生物监测平台，不仅适用于啤酒厂，也适用于其他行业。
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.