Analysis of Pork Adulteration in Commercial Burgers Targeting Porcine-Specific Mitochondrial Cytochrome B Gene by TaqMan Probe Real-Time Polymerase Chain Reaction

作者：Ali M Eaqub^*; Hashim U; Dhahi Th Sabar; Mustafa S; Man Yaakob Bin Che; Latif Md Abdul

来源：Food Analytical Methods, 2012, 5(4): 784-794.

DOI：10.1007/s12161-011-9311-4

• Burger formulation
• Double quenched TaqMan probe
• Halal and Kosher foods
• PLS model

A TaqMan probe real-time polymerase chain reaction assay was developed for the determination of pork adulteration in commercial burgers. The assay combined porcine-specific primers and TaqMan probe for the selective amplification and detection of a 109-bp fragment of swine cytochrome b (cytb) gene. Specificity test with 10 ng DNA of 11 different meat-providing animal and fish species yielded a quantification cycle (Cq) of 15.5 +/- 0.20 for the pork and negative results for the others in a 40-cycle reaction with a change of analysts and sources. Analysis of beef burger formulations with spiked pork showed the assay can determine 100-0.01% contaminated pork with a PCR efficiency (E) of 93.8% and a correlation coefficient (R (2)) of 0.991. A plot of actual value against real-time PCR-predicted value also yielded a good linear regression, R (2) 0.998, and small root mean square error of calibration, RMSEC 0.42. A strong correlation was found between the partial least square (PLS)-predicted values and real-time PCR-determined values. The accuracy of the method was a parts per thousand yen90% in all determinations of the standard set. Residual analysis also revealed a high precision in all determinations. Finally, a random analysis of 10 ng DNA of commercial burgers from pork, beef, chicken, mutton, and chevon yielded a Cq of 15.56 +/- 0.22 to 16.24 +/- 0.35 from pork burgers, and negative results from the others, showing the suitability of the assay to determine pork in commercial burgers with a high accuracy and precision.

• 出版日期2012-8