Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 69, Issue 35, Pages 10321-10328Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.1c03078
Keywords
food authentication; CRISPR-Cas12; halal food; gene analysis; Cyt b gene
Funding
- National Natural Science Foundation of China [22074100, RSP-2021/138]
- King Saud University, Riyadh, Saudi Arabia
Ask authors/readers for more resources
The study introduced a novel CRISPR-Cas12-based nucleic acid analytical strategy for the rapid identification of pork components, enhancing food authenticity verification for halal products. The method features specific discrimination of pork from other types of meat with a low detection limit.
The halal food market is globally growing along with the increased risk of adulteration. We proposed an amplification-free and mix-to-read CRISPR-Cas12-based nucleic acid analytical strategy allowing rapid identification and analysis of pork components, thus enriching the toolbox for ensuring halal food authenticity. We designed and optimized guide RNA (gRNA) targeting the pork cytochrome b (Cyt b) gene. gRNA allowed specific identification of the target Cyt b gene from pork components followed by activation of Cas12 protein to abundantly cleave single-stranded DNA probes with terminally labeled fluorophore and quencher groups, thus turning on fluorescence. The presence of the pork Cyt b gene thus can be mix-and-read- and only-one-step-detected, which may indicate the risk of halal food adulteration. The method allowed specific discrimination of pork meat from beef, mutton, and chicken and yielded a detection limit of 2.7 ng/mu L of total DNA from pork meat. The reliability of the method was tested using the following processed meat products: halal foods beef luncheon meat and spiced beef and non-halal foods sausage and dried pork slices. The CRISPR-Cas12-based nucleic acid test strategy is promising for rapid food authentication.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available