Abstract
This study aimed at evaluating the sensitivity of a multiplex PCR technique for detecting fraud by the deliberate addition of buffalo meat into the minced bovine beef. The minced bovine beef containing 0.01 %, 0.1 %, 1.0 %, 5.0 %, 10.0 %, 25.0 %, 50.0 %, 75.0 %, 90.0 %, 95.0 %, 99 %, 99.9 %, 99.99 % of minced buffalo beef were produced in triplicate, with five replicates. The minced meats exclusively of each animal species were used as control samples. Every prepared sample was analyzed by means of multiplex polymerase chain reaction. For performing the assay sensitivity analysis, the known concentrations of DNA samples (0.01 %, 0.1 %, 1.0 %, 5.0 %, 10.0 %, 25.0 %, 50.0 %, 75.0 %, 90.0 %, 95.0 %, 99.0 %, 99.9 %, 99.99 %) from Bos taurus and Bubalus bubalis species were diluted and mixed into a final volume of 10 μL; and these DNA samples were also assayed by the proposed PCR methodology. The multiplex PCR showed to be is an effective and highly sensitive technique enough for detecting increments of 10 % (2.05 ng) of bubaline DNA and of 0.1 % (0.041 ng) of bovine DNA.
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