Analysis of pork adulteration in commercial meatballs targeting porcine-specific mitochondrial cytochrome b gene by TaqMan probe real-time polymerase chain reaction |
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Authors: | Ali M E Hashim U Mustafa S Che Man Y B Dhahi Th S Kashif M Uddin Md Kamal Abd Hamid S B |
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Affiliation: | Nanotechnology and Catalysis Research Center, Universiti Malaya, 50603 Kuala Lumpur, Malaysia. eaqubali@gmail.com |
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Abstract: | A test for assessing pork adulteration in meatballs, using TaqMan probe real-time polymerase chain reaction, was developed. The assay combined porcine-specific primers and TaqMan probe for the detection of a 109 bp fragment of porcine cytochrome b gene. Specificity test with 10 ng DNA of eleven different species yielded a threshold cycle (Ct) of 15.5 ± 0.20 for the pork and negative results for the others. Analysis of beef meatballs with spiked pork showed the assay can determine 100-0.01% contaminated pork with 102% PCR efficiency, high linear regression (r(2) = 0.994) and ≤ 6% relative errors. Residuals analysis revealed a high precision in all determinations. Random analysis of commercial meatballs from pork, beef, chicken, mutton and goat, yielded a Ct between 15.89 ± 0.16 and 16.37 ± 0.22 from pork meatballs and negative results from the others, showing the suitability of the assay to determine pork in commercial meatballs with a high accuracy and precision. |
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Keywords: | Meatball formulation Double quenched TaqMan probe Halal and Kosher foods Threshold cycle Ground meat |
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