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PCR–RFLP and species-specific PCR efficiency for the identification of adulteries in meat and meat products

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Abstract

The traceability of meat origin has become a necessity to safeguard consumer's confidence in commercial meat products. Our study recommends a meat species detection using qualitative approach based on PCR–RFLP and species-specific primers PCR. Here, we targeted a 359 bp fragment of the mitochondrial cytochrome b gene amplified by PCR using universal primers followed by three enzymatic digestions. Seven animal species including dromedary, rabbit, goat, turkey, rat, donkey and pork, have been efficiently detected in pure and mixed samples. The combination of PCR–RFLP and triplex PCR assays offers, in addition, the identification of chicken, dog and cat species in meat. In conclusion, by the mean of PCR-based techniques using universal primers followed by enzymatic digestion and multiplex primer-specific approach, we developed an extensible protocol by which we identified 10 animal species that could be integrated in meat analysis daily routine.

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Data availability

All data analyzed during this study are included in this published article (and its supplementary information files).

Abbreviations

DNA:

Deoxyribonucleic acid

mtDNA:

Mitochondrial deoxyribonucleic acid

MSA:

Multiple sequence alignment

PCR:

Polymerase chain reaction

PCR–RFLP:

Restriction fragment length polymorphism polymerase chain reaction

PCR-SSCP:

Single-strand conformation polymorphism polymerase chain reaction

EDTA:

Ethylenediaminetetraacetic acid

D-loop:

Displacement loop

MgCl2 :

Magnesium chloride

dNTP:

Deoxynucleotide

DMSO:

Dimethyl sulfoxide

Cyt b:

Cytochrome b

CaSSR:

Cat species-specific region

DoSSR:

Dog species-specific region

NEBcutter:

NEB—New England Biolabs cutter

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Acknowledgements

The authors are thankful to Dr. Fatma KARRAY, Riadh BEN MARZOUG and Mohamed Arbi BEN YOUNES for providing necessary help. We also thank Prof. Saber MASMOUDI for pertinent discussions. The authors are also grateful to Kamel MAALOUL, translator and English professor for having proofread the manuscript.

Funding

The research was funded by the Ministry of Higher Education and Scientific Research of Tunisia.

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Authors and Affiliations

  1. Center of Biotechnology of Sfax, Laboratory of Molecular and Cellular Screening Processes, University of Sfax, P.O.Box ‘1177’, 3018, Sfax, Tunisia

    Héla Gargouri

  2. Private Veterinary Practice Sfax, Road Menzel Chaker km 5.5, Sfax, Tunisia

    Nizar Moalla

  3. Department of Applied Biology, College of Sciences, University of Sharjah, Office location: Building W8 - Room 116, P.O.Box: 27272, Sharjah, United Arab Emirates

    Hassen Hadj Kacem

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  1. Héla Gargouri
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Correspondence to Hassen Hadj Kacem.

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Gargouri, H., Moalla, N. & Kacem, H.H. PCR–RFLP and species-specific PCR efficiency for the identification of adulteries in meat and meat products. Eur Food Res Technol 247, 2183–2192 (2021). https://doi.org/10.1007/s00217-021-03778-y

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