数字农科院2.0

Liquid column and droplets based CRISPR/Cas12a-ultra tube microfluidic chip enabling ultra-sensitive and rapid multiplexed pathogen detection

文献类型： 外文期刊

作者： Shangyi Wu;Muzhen Wang;Zihe Xu;Fangwu Liu;Dong Dong;Baicheng Huang;Xinping Shi;Xingchi Kan;Xingxu Huang;Lina Wu;Weibo Zheng;Xuemei Tian;Xinjie Wang

作者机构：

关键词： CRISPR/Cas12a;Enteric pathogens;Microfluidic biosensor;Point-of-care multiplexed testing

期刊名称： Microchemical Journal

ISSN： 0026-265X

年卷期： 2026 年 221 卷

页码：

收录情况： SCIE（2025版）

摘要： CRISPR-based detection platforms have advanced rapid and precise nucleic acid (NA) detection for point-of-care (POC) food safety testing and disease diagnosis. However, there remains an urgent need to integrate easy-to-use target enrichment with simultaneous multitarget analysis. Herein, we introduced a novel CRISPR/AsCas12a-ultra-based Tube-integrated closed-loop microfluidic chip biosensor, termed CAT, for ultrasensitive multitarget detection. First, we developed a novel closed-loop liquid column and droplet-based ERA/CRISPR/AsCas12a-ultra system, which exhibited 3-fold intensity compared to conventional CRISPR/AsCas12a detection. Secondly, we established an automated precise liquid manipulation and real-time on-site fluorescent detection platform, enabling efficient enrichment and 8 targets parallel detection. Moreover, under optimal conditions, CAT automatically purified and detected pathogen NA targets within 35 min with the detection limit of 0.1 copy/μL(1 copy/reaction). Multiple enteric pathogens were tested simultaneously, with 100% specificity. Overall, CAT is a universal, rapid, and sensitive tube-based microfluidic platform for POC multiplexed pathogen detection.

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