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

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Abstract

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 ≥90% 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.

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Acknowledgement

M.E. Ali is a recipient of a Graduate Assistanceship from Universiti Malaysia Perlis. The project was supported by “RUGS grant no. 9031” to Prof. Y.B. Che Man and “MOSTI grant no. 05-01-35-SF-1030” to Prof. U. Hashim.

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Authors and Affiliations

  1. Institute of Nano Electronic Engineering, Universiti Malaysia Perlis, 01000, Kangar, Perlis, Malaysia

    M. Eaqub Ali, U. Hashim & Th. Sabar Dhahi

  2. Halal Products Research Institute, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia

    S. Mustafa & Yaakob Bin Che Man

  3. Department of Crop Science, Faculty of Agriculture, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia

    Md. Abdul Latif

Authors
  1. M. Eaqub Ali
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  2. U. Hashim
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  3. Th. Sabar Dhahi
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  4. S. Mustafa
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  5. Yaakob Bin Che Man
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  6. Md. Abdul Latif
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Correspondence to M. Eaqub Ali.

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Ali, M.E., Hashim, U., Dhahi, T.S. et al. Analysis of Pork Adulteration in Commercial Burgers Targeting Porcine-Specific Mitochondrial Cytochrome B Gene by TaqMan Probe Real-Time Polymerase Chain Reaction. Food Anal. Methods 5, 784–794 (2012). https://doi.org/10.1007/s12161-011-9311-4

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  • DOI: https://doi.org/10.1007/s12161-011-9311-4

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