A Sensitive Nucleic Acid Detection Platform for Foodborne Pathogens Based on CRISPR-Cas13a System Combined with Polymerase Chain Reaction

• Published in: Food analytical methods 2023-02, Vol.16 (2), p.356-366
• Main Authors: Liu, Rui, Ali, Shujat, Huang, Dong, Zhang, Yunlian, Lü, Peng, Chen, Quansheng
• Format: Article
• Language: English
• Subjects: Analytical Chemistry; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; CRISPR; Food chains; Food safety; Food Science; Foodborne diseases; Foodborne pathogens; Microbiology; Nucleic acids; Pathogens; Polymerase chain reaction; Public health; Water analysis; Water sampling
• ISSN: 1936-9751; 1936-976X
• DOI: 10.1007/s12161-022-02419-8

Foodborne diseases attributable to foodborne pathogens directly impact food safety and public health. This study aimed to establish a nucleic acid detection platform for sensitive detection of foodborne pathogens. Staphylococcus aureus (ATCC® 29213™) as a model pathogen was used to evaluate the detection performances of the developed method. The advantages of combining rapid acquisition of amplification products, crRNA programmability, and collateral cleavage effects of Cas13a were applied to achieve the detection goal. The findings implied that the target gene as low as 2.4 copies µL −1 could be detected by the method . Furthermore, compared with standard real-time PCR, the PCR-Cas13a-based nucleic acid detection method achieved satisfactory accuracy (99.44% for 180 water samples and 99.00% for 200 milk samples). With the desired simplicity, specificity, sensitivity, and accuracy, this detection strategy could be extended for the detection of other pathogens in food safety, environmental analysis, and medical diagnostics.