Authentication of caprine milk and cheese by commercial qPCR assay

Authors

  • Tomáš Fekete Slovak University of Agriculture, Faculty of Biotechnology and Food Sciences, Department of Hygiene and Food Safety, Tr. A. Hlinku 2, 949 76 Nitra
  • Marek Šnirc Slovak University of Agriculture, Faculty of Biotechnology and Food Sciences, Department of Animal Products Evaluation and Processing, Tr. A. Hlinku 2, 949 76 Nitra
  • Ľubomí­r Belej Slovak University of Agriculture, Faculty of Biotechnology and Food Sciences, Department of Hygiene and Food Safety, Tr. A. Hlinku 2, 949 76 Nitra
  • Radoslav Židek Slovak University of Agriculture, Faculty of Biotechnology and Food Sciences, Department of Hygiene and Food Safety, Tr. A. Hlinku 2, 949 76 Nitra
  • Jozef Golian Slovak University of Agriculture, Faculty of Biotechnology and Food Sciences, Department of Hygiene and Food Safety, Tr. A. Hlinku 2, 949 76 Nitra
  • Peter Haščí­k Slovak University of Agriculture, Faculty of Biotechnology and Food Sciences, Department of Animal Products Evaluation and Processing, Tr. A. Hlinku 2, 949 76 Nitra
  • Lucia Zeleňáková Slovak University of Agriculture, Faculty of Biotechnology and Food Sciences, Department of Hygiene and Food Safety, Tr. A. Hlinku 2, 949 76 Nitra Slovakia
  • Peter Zajác Slovak University of Agriculture, Faculty of Biotechnology and Food Sciences, Department of Hygiene and Food Safety, Tr. A. Hlinku 2, 949 76 Nitra

DOI:

https://doi.org/10.5219/780

Keywords:

bovine, caprine, adulteration, qPCR

Abstract

The objective of the study was to investigate potential adulteration of commercial caprine milks and cheeses with bovine milk using commercial qPCR assay. The assay comprised of bovine-, ovine- and caprine-specific primers and TaqMan probe and mammalian internal control. Specificity, sensitivity, linearity, reproducibility and efficiency of the bovine assay were tested as well. Specificity was verified by running reaction on the DNA of other milk-producing species (caprine and ovine) and made-up bovine-caprine (v/v) milk mixes. In both experiments, a bovine DNA fragment was amplified whereas no amplification was obtained from the other species. Sensitivity, linearity, reproducibility and efficiency were tested on 10-fold dilution series of 10 ng bovine DNA. The assay has shown good linearity (R2 = 0.983) within whole range, with efficiency of 86% and excellent reproducibility (SD around the CT for the technical replicates <0.5). The sensitivity was adequate, as calculated LOD and LOQ were 1.44 pg and 2.94 pg of bovine DNA, respectively. Finally, the assay was used to authenticate 5 caprine milk samples and 5 caprine cheese samples, purchased from local supermarkets. Totally, 1 milk sample has shown the fluorescence signal, which exceeded baseline in cycle 39.01 ±0.69. However, the signal was above LOD and LOQ suggesting that there could not be unambiguously declared any adulteration with bovine milk. Amplification of bovine-specific DNA was not observed in the other samples indicating products were not adulterated. The commercial qPCR assay has proved that real-time PCR assays, as well as DNA-based techniques in a general, are the excellent and reliable tools for fighting with frauds in the food industry and protecting the public health.

 

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Published

2017-10-11

How to Cite

Fekete, T. ., Šnirc, M. ., Belej, Ľubomí­r ., Židek, R. ., Golian, J. ., Haščí­k, P. ., Zeleňáková, L. ., & Zajác, P. . (2017). Authentication of caprine milk and cheese by commercial qPCR assay. Potravinarstvo Slovak Journal of Food Sciences, 11(1), 580–586. https://doi.org/10.5219/780

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