Determining the presence of chicken and turkey meat in selected meat products using realtime PCR method
Abstract
The one of the most convenient method for the identification of animal species in raw and processed meat products is the examination of DNA sequences. Real-Time PCR are particularly suitable because even small fragments of DNA formed during heat processing of the meat can be amplified and identified. TaqMan Real-Time PCR is a rapid, convenient and sensitive assay for meat identification. For chicken and turkey meat identification we were using species-specific primers and TaqMan probe designed on the mitochondrial cytochrome b. The intensity of the fluorescence signal has risen at a variety of different samples. We analysed sixteen the samples of turkey meat products and we found the incidence of chicken at nine samples in the range of the detection range of the reaction0.1 to 100%. Sample 8 fluorescence intensity exceeded the detection threshold in the 22.11 cycle (Cp = 22.11); Sample 6, (Cp = 23.19); Sample 1 in 27.08 cycle (Cp = 27.08); Sample 7 in 31,7 cycle (Cp = 31.7) and sample 5 in 32.32 cycle (Cp = 32.32). All Cp values for these samples fluorescence intensity exceeded the detection threshold in earlier cycles as sample the 100% turkey DNA. It follows that in the samples no. 8, 6, 1, 5, and 7 is in the range of chicken DNA detection range of the reaction, from 0.1 to 100%. Sample 11 in the cycle 27,08 (Cp = 27.08); Sample 10 in the cycle 27.8 (Cp = 27.8); sample 16 in 28.03 cycle (Cp = 28.03) and sample 13 in the cycle of 29.18 (Cp = 29.18). In recognition of the results of the monitoring of the content of chicken meat in meat products it is appropriate to further verification and testing detection kits used to work for possible use in practice since it has been found to be sufficient sensitivity and specificity to 30 cycle reaction.
References
Espinoza, E. O., Kirms, M. A., Filipek, M. S. 1996. Identification and quantification of source from hemoglobin of blood and blood mixtures by high performance liquid chromatography. Journal of Forensic Science. vol. 41, p. 804-811. [cit. 2014-07-15] Available at: http://www.fws.gov/lab/pdfs/espinoza_etal.1996.pdf
Hajmeer, M., Cliver, D. O., Provost, R. 2003. Spinal cord tissue detection in comminuted beef: comparison of two immunological methods. Meat Science. vol. 65, no. 2, p. 757-763. http://dx.doi.org/10.1016/S0309-1740(02)00278-4 PMid:22063437
Hird, H., Goodier, R., Hill, M. 2003. Rapid detection of chicken and turkey in heated meat products using the polymerase chain reaction followed by amplicon visualisation with vistra green. Meat Science. vol. 65, no. 3, p. 1117-1123. http://dx.doi.org/10.1016/S0309-1740(02)00341-8 PMid:22063694
Cheng, X., He, W., Huang, F., Huang, M., Zhou, G. 2014. Multiplex real-time PCR for the identification and quantification of DNA from duck, pig and chicken in Chinese blood curds. Food Research International, vol. 60, p. 30-37. http://dx.doi.org/10.1016/j.foodres.2014.01.047
Jonker, K. M., Tilburg, J. J., Hagele, G. H., De Boer, E. 2008. Species identification in meat products using real-time PCR. Food Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess. vol. 25, no. 5, p. 527-533. http://dx.doi.org/10.1080/02652030701584041 PMid:18473208
Köppel, R., Zimmerli, F., Breitenmoser, A. 2009. Heptaplex real-time PCR for the identification and quantification of DNA from beef, pork, chicken, turkey, horse meat, sheep (mutton) and goat. European Food Research and Technology, vol. 230, no. 1, p. 125-133. http://dx.doi.org/10.1007/s00217-009-1154-5
Laube, I., Zagon, J., Brpll, H. 2007. Quantitative determination of commercially relevant species in foods by real-time PCR. International Journal of Food Science & Technology, vol. 42, no. 3, p. 336-341. http://dx.doi.org/10.1111/j.1365-2621.2006.01249.x
Lepešková, I. 2002. Využití molekulárních metod při druhové identifikaci masa. Maso. vol. 5, p. 27-28.
López, I., Pardo, M. A. 2005. Application of relative quantification Taqman real-time polymerase chain reaction technology for the identification and quantification of Thunnus alalunga and Thunnus albacares. Journal of Agricultural and Food Chemistry. vol. 53, p. 4554-4560. PMid:15913324
Obrovská, I., Steinhauserová, M., Nemola, M., Krkoška, L. 2002. Identifikace druhu masa v masných výrobcích. Veterinářství., vol. 52, 2002, p. 421-423. ISSN 0506-8231.
Özgen-Arun, O., Ugur, M. 2000. Animal species determination in sausages using an SDS-PAGE technique. Archiv für Lebensmittel Hygiene. vol. 51, p. 49-53. ISSN 0003-925X.
Soares, S., Amaral, J. S., Mafra, I., Oliveira, M. B. P. P. 2010. Quantitative detection of poultry meat adulteration with pork by a duplex PCR assay. Meat Science, vol. 85, no. 3, p. 531-536. http://dx.doi.org/10.1016/j.meatsci.2010.03.001 PMid:20416827
Takáčová, D., Bugarský, A. 2010. Súdne veterinárske lekárstvo a prípady znaleckého dokazovania. Vysokoškolská učebnica. Košice : UVLF, p. 425. ISBN 978-80-8077-201-7
You, J., Huang, L., Zhuang, J., Mou, Z. 2014. Species-specific Multiplex Real-time PCR Assay for Identification of Deer and Common Domestic Animals. Food Science and Biotechnology, vol. 23, no. 1, p. 133-139. http://dx.doi.org/10.1007/s10068-014-0018-3
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).




















