Identification of Animal Species in Meat Broth by Simplex and Multiplex PCR

Peshnyar Muhammad Atta Rashid, Muhamad Omer Babashekh, Ari Salahadin Marouf, Kamaran Mohamad Amin

Molecular diagnostic laboratory, Sulaimani Veterinary directorate, Sulaimani  

In some country, some restaurants may use fraudulent substitution in meat product that has
less commercial value or undesirable. Effective laboratory tests must be available for prevention
of this phenomenon. In this study meat broth sample was successfully used to identify seven
animal species in cooked meals by simplex and multiplex PCR using species specific primers for
conserved region of mitochondrial DNA (mtDNA). The results showed effective amplification of
mtDNA without cross reaction between primers. Whereas existing methods require mincing and
homogenization of direct meat samples, this meat broth sampling method can be performed
directly and is thus easier to perform. In conclusion, meat broth sample could be used instead of
direct meat tissues for species identification of cooked meat in restaurants to control
adulteration substitutions of undesirable meat and protection of the customer rights

Keywords: Meat broth sample; species specific primer; veterinary forensic medicine; meat

[1] S. B. Ong, M. I. Zuraini, L. C. Chai, W.G. Jurin, Y. Haryani, Y. K. Cheah, F. M. Ghazali,
R. Tunung, and R. Son, "Meat Molecular Detection: Sensitivity of Polymerase Chain
Reaction-Restriction Fragment Length Polymorphism in Species Differentiation of Meat
From Animal Origin" ASEAN Food Journal 14 , pp51-59,(2007).
[2] S. Ghovvati, M. R. Nassiri, S.Z. Mirhoseini, A. HeraviMoussavi, and A. Javadmanesh,
"Fraud identification in industrial meat products by multiplex PCR assay" Food Control
20, pp696–699, (2009).
[3] A. Ciupa, M. Mihaiu, S. D. Dan, A. Lapuşan, C. Jecan, and I. Cordiş, "Using PCR
Techniques for Rapid Detection of Animal Species in Meat Products" Veterinary
Medicine 69, pp1-2, (2012).
[4] G. Arcos-Garcia, A. Totosaus, I. Guerrero, and M. L. Perez-Chabela, "Physicochemical,
sensory, functional and microbial characterisation of horse meat' Rev. Bras.
Agrociencia 8, pp43–46, (2002).
[5] O. I. Ilhak, and A. Arsalan, "Identification of Meat Species by Polymerase Chain
Reaction (PCR) Technique" Turk. J. Vet. Anim. Sci. 31, pp159-163, (2007).
[6] B. Pakendorf, and M. Stoneking, "Mitochondrial DNA and human evolution" Annual
Review of Genomics and Human Genetics 6, pp165-183, (2005).
[7] T. Matsunaga, K. Chikuni, R. Tanabe, S. Muroya, K. Shibata, J.Yamada, and Y.
Shinmura, "A quick and simple method for the identification of meat species and meat
products by PCR assay" Meat Sciences 51, pp143-148, (1999).
[8] S. Lahiff, M. O. Glennon, L. Brien, J. Lyng, T. Smith, M. Maher, and N. Shilton,
"Species-specific PCR for the identification of bovine, porcine, and chicken species in
meat and bone meal (MBM)" Mol. Cell. Probes 15, pp27-35, (2001).
[9] A. Dalmasso, E. Fontanella, P. Piatti, T. Civera, S. Rosati, and M. Bottero, "A
multiplex PCR assay for the identification of animal species in feedstuffs" Molecular and
Cellular Probes.18, pp81–87, (2004).
[10] Z. Kesmen, F. Sahin, and H. Yetim, "PCR assay for the identification of animal species
in cooked sausages" Meat Science 77, pp649-653, (2007).
[11] A. K. Winter, P. D. Thomsen, and W. Davies, "A comparison of DNA hybridization,
immunodiffusion, counter current immunoelectrophoresis and isoelectric focusing for
detecting the admixture of pork to beef" Meat Sciences 27, pp75-85, (1990).
[12] M. C. Koh, C. H. Lim, S.B. Chua, S. T. Chew, and S. T. W.Phang, "Random amplified
polymorphic DNA (RAPD) fingerprints for identification of red meat animal species"
Meat Science 48, pp275-285, (1998).
[13] D. Frezza, V. Giambra, F. Chegdani, C. Fontana, G. Maccabiani, N. Losio, E.
Faggionato, B. Chiappini, G. Vaccari, C. Holst, L. Lanni, S. Saccares, and P. Ajmone-
Marsan, "Standard and Light-Cycler PCR methods for animal DNA species detection in
animal feedstuffs" Innovative Food Science and Emerging Technologies 9, pp18-23,