Integrated slip valve-assisted fluidic chip coupling with CRISPR/Cas12a system for nucleic acid analysis
文献类型： 外文期刊
作者： Qian, Siwenjie 1 ; Chen, Yanju 1 ; Peng, Cheng 2 ; Wang, Xiaofu 2 ; Che, Yang 3 ; Wang, Tingzhang 3 ; Wu, Jian 1 ; Xu, Junfeng 2 ;
作者机构： 1.Zhejiang Univ, Coll Biosyst Engn & Food Sci, Hangzhou 310058, Peoples R China
2.Zhejiang Acad Agr Sci, Key Lab Traceabil Agr Genet Modified Organisms, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China
3.Zhejiang Inst Microbiol, Key Lab Microbial Technol & Bioinformat Zhejiang P, Hangzhou 310012, Peoples R China
4.Minist Agr & Rural Affairs, Key Lab Site Proc Equipment Agr Prod, Hangzhou 310058, Peoples R China
关键词： Nucleic acid detection; Integrated chip; Dual detection; Loop -mediated isothermal amplification; CRISPR; Cas12a
期刊名称：ANALYTICA CHIMICA ACTA （ 影响因子：6.2； 五年影响因子：5.9 ）
ISSN： 0003-2670
年卷期： 2023 年 1239 卷
页码：
收录情况： SCI
摘要： Currently, some on-site nucleic acid detection platforms have been developed. However, these platforms still need to be improved in device integration and multiple detection capability. In this work, an integrated dual nucleic acid analysis platform was developed by slip valve-assisted fluidic chip coupled with CRISPR/Cas12a system. All the reagents, including nucleic acid extraction, air-dried loop-mediated isothermal amplification (LAMP) and CRISPR/Cas12a detection reagents, were preloaded on the fluidic chip. Liquids transfer and stirring could be controlled by a slip valve and a syringe. By combining duplex LAMP reaction with two CRISPR detection units, CRISPR/Cas12a-based dual nucleic acid analysis was successfully constructed. Benefiting from highquality nucleic acid extraction on the chip, as low as 30 copies/reaction of Vibrio parahaemolyticus (V. parahaemolyticus) and 20 copies/reaction of Salmonella typhimurium (S. typhimurium) could be simultaneously detected. Detection results could be observed by the naked eye under a portable ultraviolet lamp. The whole
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