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

Meat Sci. 2012 Aug;91(4):454-9. doi: 10.1016/j.meatsci.2012.02.031. Epub 2012 Mar 6.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Animals
  • Chickens / genetics
  • Cytochromes b / genetics*
  • Genes, Mitochondrial*
  • Livestock / genetics
  • Meat Products / analysis*
  • Meat Products / standards
  • Mitochondria / genetics*
  • Real-Time Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Swine / genetics*

Substances

  • Cytochromes b