CRISPR-Cas12-Based Rapid Authentication of Halal Food.
J Agric Food Chem
; 69(35): 10321-10328, 2021 Sep 08.
Article
in En
| MEDLINE
| ID: mdl-34436881
ABSTRACT
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/µL 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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Red Meat
/
Meat Products
Type of study:
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
J Agric Food Chem
Year:
2021
Document type:
Article
Affiliation country:
China
Publication country:
EEUU
/
ESTADOS UNIDOS
/
ESTADOS UNIDOS DA AMERICA
/
EUA
/
UNITED STATES
/
UNITED STATES OF AMERICA
/
US
/
USA