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 degrees 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/mu L 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.