Multiplex bacteria detection using one-pot CRISPR_Cas13a尚玉婷

High-throughput detection of bacteria at low levels is critical in public health, food safety, and first response.Herein, for the first time, we present a platfomm based on droplet microfluidics coupling with the recombinaseaided amplification (R AA)-assisted one-pot clustered regularly interspaced short palindro mic repeats togetherwith CRlSPR-associated proteins 13a (CRlSPR/Cas13a) asay, and dro plet encoding strategy for accurate andsensitive determination of nucleic acids from various foodborne pat hogens., "'he workilow takes full advantage ofCRISPR/Casl 3a signal amplification and dro plet confinement effects, which enhances the detection sensitivityand enables end- point quantitat lon, Meanwhile, by varying the color of dro plets, the number of bacteria detectedat the same time is greatly improved. It possesses the capability to simultaneously detect seven different ty pes offoodborne pat hogens, Notably, the system is also applied to real food samples with satisfactory results. Overallin view of superorities in high sensitivity, outstanding selectivity, and large-scale multiplexing, the one- potCRlSPR/Casl 3a-based droplet microlluidic system could be expanded and universalized for identifying otherbactena.

文件下载