Multi-throughput POCT Technology Based on RPA and CRISPR/Cas12a and Its Application in Detection of Adulterated Meat

Mixing low-priced meat (pork, chicken, and duck) into high-priced meat (beef and mutton) not only deceives consumers but also involves issues of food safety and religious beliefs. In this study, an on-site gene detection technology was established by universal primer-mediated recombinase polymerase amplification (RPA) combined with a specific CRISPR/Cas12a system, which was applied to nondirectional detection in common adulterated meat products. Within one hour at 37 °C, this technology can specifically be used to identify the meat source of beef, mutton, pork, chicken, and duck through fluorescent signals. The detection sensitivity is up to 10 pg/μL DNA concentration, and 1% incorporation of low-priced meat sources in high-priced meat sources could be detected by this technology. Through the integration of RPA and CRISPR system with microfluidic equipment, the on-site point-of-care testing is realized, which can be applied for the specific, sensitive, rapid, and multiple identification of adulterated meat.