Determination of the species specificity of the primers for the detection of chicken and turkey meat by realtime PCR method

Authors

  • Lenka Maršálková Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of Food Hygiene and Safety, Tr. A. Hlinku 2, 949 76 Nitra
  • Miloš Mašlej Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of Food Hygiene and Safety, Tr. A. Hlinku 2, 949 76 Nitra
  • Ľubomí­r Belej Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of Food Hygiene and Safety, Tr. A. Hlinku 2, 949 76 Nitra
  • Jozef Golian Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of Food Hygiene and Safety, Tr. A. Hlinku 2, 949 76 Nitra
  • Radoslav Židek Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of Food Hygiene and Safety, Tr. A. Hlinku 2, 949 76 Nitra

DOI:

https://doi.org/10.5219/390

Keywords:

primer, chicken meat, turkey meat, PCR, TaqMan

Abstract

The aim of this work was to use TaqMan Real-Time PCR for quantitative authentication of chicken and turkey meat.
To meet this purpose, a specific pair of primers and TaqMan probe was used. The test was aimed at identifying the reaction cycle of turkey and chicken meat using by two sets of primers. With first set of primer designed for chicken we obtained the following results: Cp = 16.18 for 100% chicken DNA Cp = 29, 18 100% turkey DNA It was also amplified DNA of pig that exceeded the detection threshold fluorescence intensities in the 31.07 cycle (Cp = 31.07). Using primers designed for turkey we obtained the following results Cp = 31.16 for 100% CHDNA, Cp =16.18 100% TDNA. It was also amplified the 100% DNA of rabbit in 31.63 cycle (Cp = 31.63) and deer in cycle 32 (Cp = 32). The DNA of all other animal species was amplificated after more than 35 cycles (Cp >35). It follows that the second detection primer pair is specific enough to unrelated species of animals by 30 cycles of the reaction. Species authentication based on DNA analysis from this perspective overcomes all the shortcomings of proteins. At present, DNA analysis use different types of PCR. Is the most progressive Real-time PCR, which is suitable for the specific use of detection (primers and TaqMan probe). The TaqMan Real-time PCR is within the sensitivity and specificity, clearly one of the best methods for identifying the species of chicken and turkey meat. The specificity of this method, however, depends primarily on the specificity of the primers and TaqMan probe. The 30 cycle reaction was chosen by us as the threshold for specificity using primers for authentication chicken and turkey meat.

Downloads

Download data is not yet available.

Metrics

Metrics Loading ...

References

Cammà, C., Di Domenico, M., Monaco, F. 2012. Development and validation of fast Real-Time PCR assays for species identification in raw and cooked meat mixtures. Food Control, vol. 23, no. 12, p. 400-404. https://doi.org/10.1016/j.foodcont.2011.08.007 DOI: https://doi.org/10.1016/j.foodcont.2011.08.007

Ciglenečky, U. J., Grom, J., Toplak, I., Jemeršić, L., Barlič-Maganja, D. 2008. Real-time RT-PCR assay for rapid and specific detection of classical swine fever virus: Comparison of SYBR Green and TaqMan MGB detection methods using novel MGB probes. Journal of Virological Methods, vol. 14, no. 2, p. 257-264. https://doi.org/10.1016/j.jviromet.2007.09.017 DOI: https://doi.org/10.1016/j.jviromet.2007.09.017

Dooley, J. J, Paine, K. E., Garrett, S. D., Brown, H. M. 2004. Detection of meat species using TaqMan real-time PCR assays. Meat Science, vol. 68, no. 3, p. 431-438. https://doi.org/10.1016/j.meatsci.2004.04.010 PMid:22062411 DOI: https://doi.org/10.1016/j.meatsci.2004.04.010

Jonker, K. M., Tilburg, J. J., Hägele, G. H., DeBoer, 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. https://doi.org/10.1080/02652030701584041 PMid:18473208 DOI: https://doi.org/10.1080/02652030701584041

Haider, N., Nabulsi, I., Al-Safadi, B. 2012. Identification of meat species by PCR-RFLP of the mitochondrial COI gene. Meat Science, vol. 90, no. 2, p. 490-493. https://doi.org/10.1016/j.meatsci.2011.09.013 PMid:21996288 DOI: https://doi.org/10.1016/j.meatsci.2011.09.013

Kesmen, Z., Yetiman, A. E., Sahin, F., Yetim, H. 2012. Detection of chicken and turkey meat in meat mixtures by using Real-time PCR assays. Journal of Food Science, vol. 77, no. 2, p. C167-C173. https://doi.org/10.1111/j.1750-3841.2011.02536.x PMid:22309374 DOI: https://doi.org/10.1111/j.1750-3841.2011.02536.x

Králová, B., Fukal, L., Rauch, P., Ruml, T. 2007. Bioanalytické metody. 3. Ed. Praha: VŠCHT, 254 p. ISBN 978-807080-449-3.

Laube, I., Zagon, J., Broll, 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. https://doi.org/10.1111/j.1365-2621.2006.01249.x DOI: https://doi.org/10.1111/j.1365-2621.2006.01249.x

Lees, M. 2003. Food autenticity and traceability. Woodhead Publishig Limited, 400 p. ISBN 978-1-85573-526-2.

Peris, M., Escuder-Gilabert, L. 2009. A 21 st century technice for food control: electronic noses. Analytica Chimica Acta. vol. 638, no. 1, p. 1-15. https://doi.org/10.1016/j.aca.2009.02.009 PMid:19298873 DOI: https://doi.org/10.1016/j.aca.2009.02.009

Omelka, R., Bauerová, M., Laurinčík, I. 2001. Optimalizácia metódy PCR-RFLP na detekciu Pvu II polymorfizmu estrogénového repertoáru ošípaných. Zborník z II. vedeckej konferencie doktorandov FPV s medzinárodnou účasťou. Nitra: FPV UKF, p. 279-283. ISBN-80-8050-386-9.

Popelka, P., Horská, D., Golian, J. Marcinčák, S. 2002. Detekcia falšovania ovčieho mlieka a syrov pomocou enzýmovej imunoanalýzy (ELISA). Slovenský veterinársky časopis. ISSN 1335-0099, vol. 27, no. 3, p. 36-37.

Rojas, M., Gonzáles, I., García, T., Hernández, P. E., Martín, R. 2012. Authentication of meat and commercial meat products from common pigeon (Columba livia) woodpigeon (Columba palumbus) and stock pigeon (Columba oenas) using a TaqMan® real-time PCR assay. Food Control, vol. 23, no. 2, p. 369-376. ISSN 0956-7135. DOI: https://doi.org/10.1016/j.foodcont.2011.07.034

Šmarda, J. et al. 2008. Metody molekulární biologie. Brno-Kraví Hora: Masarykova univerzita, 188 p. ISBN 978-80-210-3841-7.

Španová, A., Rittich, B., Beneš, M., Horák, D. 2005. Ferrite suports for isolation of DNA from complex samples and polymerase chain reaction amplification. Journal of Chromatography. ISSN 0021-9673, 1080, p 93-98. DOI: https://doi.org/10.1016/j.chroma.2005.05.006

Takačová, D., Bugarský, A. 2010. Súdne veterinárske lekárstvo a prípady znaleckého dokazovania. Vysokoškolská učebnica. Košice: UVLF, 425 p. ISBN 978-80-8077-201-7.

Tanabe, S., Hase, M., Yano, T., Sato, M., Fujimura, T., Akiyama, H. 2007. A real-time quantitative PCR detection method for pork, chicken, beef, mutton, and horseflesh in foods. Bioscience, Biotechnology, and Biochemistry, vol. 71, no. 12, p. 3131-3135. PMid:18071237 DOI: https://doi.org/10.1271/bbb.70683

Yilmaz, A., Onen, H. I., Alp, E., Menevse, S. 2012. Real-Time PCR for Gene Expression Analysis. Polymerase Chain Reaction, Dr Patricia Hernandez-Rodriguez (Ed.), ISBN: 978-953-51-0612-8, [cit. 2014-07-10] Available at: http://cdn.intechopen.com/pdfs-wm/37270.pdf DOI: https://doi.org/10.5772/37356

Downloads

Published

2014-07-21

How to Cite

Maršálková, L. ., Mašlej, M. ., Belej, Ľubomí­r ., Golian, J. ., & Židek, R. . (2014). Determination of the species specificity of the primers for the detection of chicken and turkey meat by realtime PCR method. Potravinarstvo Slovak Journal of Food Sciences, 8(1), 216–220. https://doi.org/10.5219/390

Similar Articles

You may also start an advanced similarity search for this article.