Direct lysis-multiplex PCR assay for detecting pork, chicken, and duck in beef products

The assay was created in order to provide a cheaper, easier, and faster method of detecting meat species substitution. A new direct lysis (DL) approach was utilized to extract DNA directly from meat tissue and get a sample of DNA for multiplex PCR in less than 15 minutes, which is significantly faster than using a commercial lysis kit. For the mitochondrial D-loop region of beef, pork, chicken, and duck, four pairs of high-specificity primers were constructed. Even after freezing, heating, and autoclaving, the assay could detect pork, chicken, and duck down to 0.1 % (w/w) in meat mixes. The assay was tested on 79 commercial beef items obtained from local markets, and it was discovered that roughly 28% of them contained pork or chicken in some form. By repeating the study using a conventional real-time PCR, the correctness of the data was confirmed.

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https://www.sciencedirect.com/science/article/abs/pii/S095671352100390X