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 high-quality 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 detection procedure was finished within 60 min. This method with integrated nucleic acid analysis, dual detection capability and fluorescence visualized results provides a new solution for on-site nucleic acid analysis.

目前，已经开发了一些现场核酸检测平台。然而，这些平台在设备集成和多重检测能力方面仍有待提高。在这项工作中，通过滑阀辅助流控芯片与 CRISPR/Cas12a 系统耦合开发了一个集成的双核酸分析平台。所有试剂，包括核酸提取、风干环介导等温扩增 (LAMP) 和 CRISPR/Cas12a 检测试剂，都预装在流控芯片上。液体转移和搅拌可以通过滑阀和注射器来控制。通过将双重 LAMP 反应与两个 CRISPR 检测单元相结合，成功构建了基于 CRISPR/Cas12a 的双重核酸分析。受益于芯片上的高质量核酸提取，低至 30 拷贝/反应副溶血性弧菌（V. parahaemolyticus）和20拷贝/反应的鼠伤寒沙门氏菌（S. typhimurium）可同时检测。检测结果可在便携式紫外灯下用肉眼观察。整个检测过程在60分钟内完成。这种集核酸分析、双重检测能力和荧光可视化结果于一身的方法为现场核酸分析提供了一种新的解决方案。