Analysis of pork adulteration in commercial meatballs targeting porcine-specific mitochondrial cytochrome b gene by TaqMan probe real-time polymerase chain reaction

• DOI: 10.1016/j.meatsci.2012.02.031
• Publication Date: 2012-03-06T20:46:51Z
• Authors: M.E. Ali; U. Hashim; S. Mustafa; Y.B. Che Man; Th.S. Dhahi; M. Kashif; Md. Kamal Uddin; S.B. Abd Hamid
• Keywords: Livestock; Swine; Reproducibility of Results; 04 agricultural and veterinary sciences; Cytochromes b; Real-Time Polymerase Chain Reaction; Mitochondria; 3. Good health; Meat Products; Genes, Mitochondrial; Animals; 0405 other agricultural sciences; Chickens

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.