Advanced SearchSearch Tips
Meat Species Identification using Loop-mediated Isothermal Amplification Assay Targeting Species-specific Mitochondrial DNA
facebook(new window)  Pirnt(new window) E-mail(new window) Excel Download
 Title & Authors
Meat Species Identification using Loop-mediated Isothermal Amplification Assay Targeting Species-specific Mitochondrial DNA
Cho, Ae-Ri; Dong, Hee-Jin; Cho, Seongbeom;
  PDF(new window)
Meat source fraud and adulteration scandals have led to consumer demands for accurate meat identification methods. Nucleotide amplification assays have been proposed as an alternative method to protein-based assays for meat identification. In this study, we designed Loop-mediated isothermal amplification (LAMP) assays targeting species-specific mitochondrial DNA to identify and discriminate eight meat species; cattle, pig, horse, goat, sheep, chicken, duck, and turkey. The LAMP primer sets were designed and the target genes were discriminated according to their unique annealing temperature generated by annealing curve analysis. Their unique annealing temperatures were found to be for cattle, for pig, and for horse in the BSE-LAMP set (Bos taurus, Sus scrofa domesticus and Equus caballus); for goat and for sheep in the CO-LAMP set (Capra hircus and Ovis aries); and for chicken, for duck, and for turkey in the GAM-LAMP set (Gallus gallus, Anas platyrhynchos and Meleagris gallopavo). No cross-reactivity was observed in each set. The limits of detection (LODs) of the LAMP assays in raw and cooked meat were determined from to levels, and LODs in raw and cooked meat admixtures were determined from 0.01% to 0.0001% levels. The assays were performed within 30 min and showed greater sensitivity than that of the PCR assays. These novel LAMP assays provide a simple, rapid, accurate, and sensitive technology for discrimination of eight meat species.
LAMP;meat identification;mitochondrial DNA;meat adulteration;food safety;
 Cited by
Application of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cow Components Adulterated in Buffalo Milk/Meat, Molecular Biotechnology, 2016, 58, 12, 850  crossref(new windwow)
A Comparison and Integration of MiSeq and MinION Platforms for Sequencing Single Source and Mixed Mitochondrial Genomes, PLOS ONE, 2016, 11, 12, e0167600  crossref(new windwow)
DNA-based techniques for authentication of processed food and food supplements, Food Chemistry, 2018, 240, 767  crossref(new windwow)
Ahmed, M. U., Hasan, Q., Mosharraf Hossain, M., Saito, M., and Tamiya, E. (2010) Meat species identification based on the loop mediated isothermal amplification and electrochemical DNA sensor. Food Control 21, 599-605. crossref(new window)

Avise, J. C., Arnold, J., Ball, R. M., Bermingham, E., Lamb, T., Neigel, J. E., Reeb, C. A., and Saunders, N. C. (1987) Intraspecific phylogeography-the mitochondrial-DNA bridge between population-genetics and systematics. Annu. Rev. Ecol. Syst. 18, 489-522. crossref(new window)

Ballin, N. Z., Vogensen, F. K., and Karlsson, A. H. (2009) Species determination-Can we detect and quantify meat adulteration? Meat Sci. 83, 165-174. crossref(new window)

Bottero, M. T. and Dalmasso, A. (2011) Animal species identification in food products: evolution of biomolecular methods. Vet. J. 190, 34-38. crossref(new window)

Cheng, Y. H., Cheng, S. D., and Weng, C. F. (2006) Investigation of goats' milk adulteration with cows' milk by PCR. Asian Australas. J. Anim. Sci. 19, 1503-1507. crossref(new window)

Chikuni, K., Ozutsumi, K., Koishikawa, T., and Kato, S. (1990) Species identification of cooked meats by DNA hybridization assay. Meat Sci. 27, 119-128. crossref(new window)

Chisholm, J., Conyers, C., Booth, C., Lawley, W., and Hird, H. (2005) The detection of horse and donkey using real-time PCR. Meat Sci. 70, 727-732. crossref(new window)

Dalmasso, A., Fontanella, E., Piatti, P., Civera, T., Rosati, S., and Bottero, M. T. (2004) A multiplex PCR assay for the identification of animal species in feedstuffs. Mol. Cell Probes. 18, 81-87. crossref(new window)

Dawnay, N., Ogden, R., McEwing, R., Carvalho, G. R., and Thorpe, R. S. (2007) Validation of the barcoding gene COI for use in forensic genetic species identification. Forensic Sci. Int. 173, 1-6. crossref(new window)

Dincer, B., Spearow, J. L., Cassens, R. G. and Greaser, M. L. (1987) The effects of curing and cooking on the detection of species origin of meat products by competitive and indirect ELISA techniques. Meat Sci. 20, 253-265. crossref(new window)

Dooley, J. J., Paine, K. E., Garrett, S. D., and Brown, H. M. (2004) Detection of meat species using TaqMan real-time PCR assays. Meat Sci. 68, 431-438. crossref(new window)

Francois, P., Tangomo, M., Hibbs, J., Bonetti, E. J., Boehme, C. C., Notomi, T., Perkins, M. D., and Schrenzel, J. (2011) Robustness of a loop-mediated isothermal amplification reaction for diagnostic applications. FEMS Immunol. Med. Microbiol. 62, 41-48. crossref(new window)

Gill, C. O. (2005) Safety and storage stability of horse meat for human consumption. Meat Sci. 71, 506-513. crossref(new window)

Girish, P. S., Anjaneyulu, A. S. R., Viswas, K. N., Anand, M., Rajkumar, N., Shivakumar, B. M., and Bhaskar, S. (2004) Sequence analysis of mitochondrial 12S rRNA gene can identify meat species. Meat Sci. 66, 551-556. crossref(new window)

Hebert, P. D., Stoeckle, M. Y., Zemlak, T. S., and Francis, C. M. (2004) Identification of birds through DNA barcodes. PLoS Biology 2, e312. crossref(new window)

Hitchcock, C. H. and Crimes, A. A. (1985) Methodology for meat species identification: A review. Meat Sci. 15, 215-224. crossref(new window)

Kesmen, Z., Gulluce, A., Sahin, F., and Yetim, H. (2009) Identification of meat species by TaqMan-based real-time PCR assay. Meat Sci. 82, 444-449. crossref(new window)

Kesmen, Z., Yetiman, A. E., Sahin, F., and Yetim, H. (2012) Detection of chicken and turkey meat in meat mixtures by using real-time PCR assays. J. Food Sci. 77, C167-173. crossref(new window)

Koh, B. R. D., Kim, J. Y., Na, H. M., Park, S. D., and Kim, Y. H. (2011) Development of species-specific multiplex PCR assays of mitochondrial 12S rRNA and 16S rRNA for the identification of animal species. Korean J. Vet. Serv. 34, 417-428. crossref(new window)

Macedo-Silva, A., Barbosa, S. F., Alkmin, M. G., Vaz, A. J., Shimokomaki, M., and Tenuta-Filho, A. (2000) Hamburger meat identification by dot-ELISA. Meat Sci. 56, 189-192. crossref(new window)

Martin, I., Garcia, T., Fajardo, V., Lopez-Calleja, I., Hernandez, P. E., Gonzalez, I., and Martin, R. (2007) Species-specific PCR for the identification of ruminant species in feedstuffs. Meat Sci. 75, 120-127. crossref(new window)

Martinez, I. and Malmheden Yman, I. (1998) Species identification in meat products by RAPD analysis. Food Res. Int. 31, 459-466. crossref(new window)

Mouillesseaux, K. P., Klimpel, K. R., and Dhar, A. K. (2003) Improvement in the specificity and sensitivity of detection for the Taura syndrome virus and yellow head virus of penaeid shrimp by increasing the amplicon size in SYBR Green real-time RT-PCR. J. Virol. Methods 111, 121-127. crossref(new window)

Pappas, C., Tarantilis, P., Moschopoulou, E., Moatsou, G., Kandarakis, I., and Polissiou, M. (2008) Identification and differentiation of goat and sheep milk based on diffuse reflectance infrared fourier transform spectroscopy (DRIFTS) using cluster analysis. Food Chem. 106, 1271-1277. crossref(new window)

Proom, H. (1943) The preparation of precipitating sera for the identification of animal species. J. Pathol. Bacteriol. 55, 419-426. crossref(new window)

Rodriguez, M. A., Garcia, T., Gonzalez, I., Asensio, L., Mayoral, B., Lopez-Calleja, I., Hernandez, P. E., and Martin, R. (2003) Identification of goose, mule duck, chicken, turkey, and swine in foie gras by species-specific polymerase chain reaction. J. Agr. Food Chem. 51, 1524-1529. crossref(new window)

Sakalar, E. and Abasiyanik, M. F. (2012) The devolopment of duplex real-time PCR based on SYBR Green florescence for rapid dentification of ruminant and poultry origins in foodstuff. Food Chem. 130, 1050-1054. crossref(new window)

Santos, C. G., Melo, V. S., Amaral, J. S., Estevinho, L., Oliveira, M. B., and Mafra, I. (2012) Identification of hare meat by a species-specific marker of mitochondrial origin. Meat Sci. 90, 836-841. crossref(new window)

Tomita, N., Mori, Y., Kanda, H., and Notomi, T. (2008) Loopmediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nat. Protoc. 3, 877-882. crossref(new window)

Varga, A. and James, D. (2005) Detection and differentiation of Plum pox virus using real-time multiplex PCR with SYBR Green and melting curve analysis: a rapid method for strain typing. J. Virol. Methods 123, 213-220. crossref(new window)

Verkaar, E. L. C., Nijman, I. J., Boutaga, K., and Lenstra, J. A. (2002) Differentiation of cattle species in beef by PCRRFLP of mitochondrial and satellite DNA. Meat Sci. 60, 365-369. crossref(new window)

Whittaker, R. G., Spencer, T. L., and Copland, J. (1983) An enzyme-linked immunosorbent assay for species identification of raw meat. J. Sci. Food Agr. 34, 1143-1148. crossref(new window)

Wolf, C., Rentsch, J., and Hubner, P. (1999) PCR-RFLP analysis of mitochondrial DNA: a reliable method for species identification. J. Agr. Food Chem. 47, 1350-1355. crossref(new window)

You, J., Huang, L., Zhuang, J., and Mou, Z. (2014) Speciesspecific multiplex real-time PCR assay for identification of deer and common domestic animals. Food Sci. Biotechnol. 23, 133-139. crossref(new window)

Zhang, C. L., Fowler, M. R., Scott, N. W., Lawson, G., and Slater, A. (2007) A TaqMan real-time PCR system for the identification and quantification of bovine DNA in meats, milks and cheeses. Food Control 18, 1149-1158. crossref(new window)