Application of Visual Saltatory Rolling Circle Amplification Method in Detection of Adulterated Pork

Objective:A saltatory rolling circle amplification(SRCA)combined with the fluorescent dye SYBR Green I was developed to detect the adulteration of pork in meat products. Methods:According to the DNA sequences of pork from NCBI database,pork-specifie primers for conventional SRCA and fluorescence visualization SRCA were designed by DNAMAN software. Fresh muscle tissue samples of nine different species were used to confirm the specificity of SRCA. Moreover,66 commercial meat products with unlabeled pork ingredients were tested for pork ingredients. Conclusion:The detection sensitivity of SRCA method for pork DNA was 6.5 fg/L by visualization with fluorescence,with conventional PCR method,the SRCA achieved 1000-fold higher sensitivity. In artificially added pork samples,the minimum detectable proportion of pork component in food samples was 0.01%(w/w). The results were 100% sensitivity,96.66% specificity and 98.48% accuracy compared with the industry standard NY/T 3309-2018. Conclusion:SRCA would be a sensitive and visual detection method for the detection of pork adulteration in meat products with great application potential.