Swine-Specific PCR-RFLP Assay Targeting Mitochondrial Cytochrome B Gene for Semiquantitative Detection of Pork in Commercial Meat Products |
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Authors: | M Eaqub Ali U Hashim S Mustafa and Yaakob Bin Che Man |
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Affiliation: | (1) Institute of Nano Electronic Engineering (INEE), Universiti Malaysia Perlis, 01000 Kangar, Perlis, Malaysia;(2) Halal Products Research Institute, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia; |
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Abstract: | Verification of pork adulteration in commercial meat products is increasingly important for the authentication of Halal labels
in processed foods. Here, we documented a PCR–restriction fragment length polymorphism (RFLP) assay with high precision and
reproducibility for the tracing of porcine DNA in commercial meat products. The assay combined the species-specific primers
to selectively amplify a short fragment (109 bp) of porcine cytochrome b gene from a heterogeneous background of genomic DNAs
followed by RFLP analysis to authenticate real amplicon. The analysis of PCR products and restriction digests was automated
in a chip-based capillary electrophoresis incorporated in Agilent 2100 bioanalyzer. The swine specificity of the assay was
checked with 11 different meat-providing animal and fish species. Model experiments, mimicking the processed foods, were performed
in binary and ternary mixtures after mechanical grinding and prolonged autoclaving. Finally, four types of the most popular
finished meat products (meatball, streaky beacon, frankfurter, and burger) which are prevalent in the Malaysian food market
were analyzed in order to verify the assay performance. The assay was sensitive enough to detect 0.0001 ng of swine DNA in
pure formats and 0.01% (w/w) spiked pork in extensively processed ternary mixture of pork, beef, and wheat flour. |
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