肉类真实性对于保障食品安全及消费者权益意义重大，掺假检测技术是保障肉类真实性的重要支撑。现有肉类掺假检测技术主要以大型仪器为主，便携检测技术发展缓慢，不能满足现场检测需求。本项目结合申请人前期研究基础，拟建立基于CRISPR/Cas9系统和表面增强拉曼技术的便携、灵敏和多重肉类真实性检测平台。利用RPA等温扩增实现掺假肉类DNA指纹便携扩增，设计掺假肉类mRNA激活的sgRNA“开关”引导CRISPR/Cas9特异性剪切标记拉曼标签的桥连DNA，通过拉曼检测将掺假肉类信息转化为拉曼信号输出；构建不同掺假肉类对应的sgRNA“开关”和SERS标签，根据拉曼信号响应差异实现多重检测。凭借CRISPR/Cas9在sgRNA引导下特异性DNA剪切功能，SERS报告物低生物背景等优势，实现高灵敏多重肉类掺假便携检测。本项目将为肉类真实性监管提供关键检测技术，为了解我国肉类掺假现状提供基础数据。

Meat authenticity is of great significance to ensure food safety and consumers' interests. Adulteration detection technology is an important support for meat authenticity. Current methodologies mainly depend on sophisticated equipment and portable detection technology develop slowly, which cannot meet the demands of point-of-care testing. Based on the research background, this project intends to establish a portable, sensitive and multiple meat authenticity detection platform via CRISPR/Cas9 and surface enhanced Raman scattering. The information of meat adulteration will be converted into Raman signal output through RPA isothermal amplification to achieve portable amplification of adulterated meat DNA fingerprints, and sgRNA "switches" activated by adulterated meat mRNA to guide CRISPR/Cas9 to specifically cleave bridged DNA labeled with Raman tags. Moreover, sgRNA "switches" and SERS tags corresponding to different adulterated meats are constructed to realize multiple detection based on different response of Raman signals. Relying on the advantages of specific DNA cleavage of CRISPR/Cas9 under the guidance of sgRNA and SERS reporters with low biological background, portable and sensitive detection of multiple meat adulteration will be achieved. This project will provide key technologies for the supervision of meat authenticity and basic data for understanding the status of meat adulteration in China.