Comparison of PCR Methods for Determination of Different Types of Milk Added to Goat Milk

Yıl 2022, Cilt: 11 Sayı: 3, 509 - 514, 04.10.2022

Rabia Mehtap Tuncay , Yakup Can Sancak

https://doi.org/10.53424/balikesirsbd.1139179

Cited By: 1

Öz

Aim: In this study, it was aimed to determine which of the multiplex conventional PCR and Real-Time PCR methods are more suitable for the detection of cow and sheep milk mixed with goat milk. Materials and Methods: For this purpose, one liter of each goat, cow, and sheep milk was obtained from farms in Van province. PCR experiments were carried out by adding cow's milk and sheep's milk in the same proportions into goat milk (1%, 2%, 5%, 0.1%, and 0.5%). Multiplex Conventional and Real-Time PCR were used in these trials. Results: In the cow and sheep milk trials, it was determined that the presence of 1%, 2%, and 5% cow and sheep milk added to goat milk could be determined by the multiplex conventional PCR method. However, it was observed that the positivity of the gel image of the milk mixtures added at the rate of 0.5% was unclear, and the mixtures at the rate of 0.1% could not be detected. In the Real-Time PCR method, the presence of cow and sheep milk was detected in all the mixtures and positive graphics were determined. Conclusion: This showed that the Real-Time PCR method gives more reliable results even when 0.1% cow or sheep milk is mixed with commercially available goat milk.

Anahtar Kelimeler

Goat milk, Multiplex conventional PCR, Real Time PCR

Destekleyen Kurum

Van Yüzüncü Yıl Üniversitesi

Proje Numarası

TSA-2020-8901

Teşekkür

This study was supported by the Research Fund of Van YuzuncuYıl University with the project number of TSA-2020-8901. Thank you for your contributions. We would like to thank Tuncer Çakmak for his support.

Keçi Sütüne Eklenen Farklı Süt Türlerinin Belirlenmesi İçin PCR Yöntemlerinin Karşılaştırılması

Öz

Amaç: Bu çalışma, keçi sütüne karıştırılmış inek ve koyun sütünün tespiti için multipleks konvansiyonel PCR ve Real Time PCR yöntemlerinden hangisinin daha uygun olduğunun belirlenmesi amavıyla yapılmıştır. Gereç ve Yöntem: Bu amaçla Van ilindeki çiftliklerden keçi, inek ve koyun sütünden birer litre süt temin edilmiştir. Keçi sütüne aynı oranlarda inek sütü ve koyun sütü (%1, %2, %5, %0.1 ve %0.5) ilave edilerek PCR deneyleri yapılmıştır. Bu denemelerde mültipleks konvansiyonel ve Real Time PCR kullanıldı. Bulgular: İnek ve koyun sütü denemelerinde keçi sütüne ilave edilen %1, %2 ve %5 inek ve koyun sütünün varlığının multipleks konvansiyonel PCR yöntemi ile belirlenebileceği ortaya konmuştur. Ancak %0.5 oranında eklenen süt karışımlarının jel görüntüsünün pozitifliğinin belirsiz olduğu ve %0.1 oranındaki karışımların tespit edilemediği görülmüştür. Real Time PCR yönteminde ise tüm karışımlarda inek ve koyun sütü varlığı tespit edildi ve pozitif grafikler belirlendi. Sonuç: Bu sonuçlar, Real Time PCR yönteminin, ticari olarak satılan keçi sütü ile %0,1 inek veya koyun sütü karıştırıldığında bile daha güvenilir sonuçlar verdiğini gösterdi.

Anahtar Kelimeler

Keçi sütü, Multiplex konvansiyonel PCR, Real Time PCR

Proje Numarası

TSA-2020-8901

Kaynakça

• Abbas, O., Zadravec, M., Baeten, V., Mikuš, T., Lešić, T., Vulić, A., Prpić, J., Jemeršić, L., & Pleadin, J. (2018). Analytical methods used for the authentication of food of animal origin. Food Chemistry, 246, 6–17. https://doi.org/10.1016/j.foodchem.2017.11.007
• Agrimonti, C., Pirondini, A., Marmiroli, M., & Marmiroli, N. (2015). A quadruplex PCR (qxPCR) assay for adulteration in dairy products. Food Chemistry, 187, 58-64. https://doi.org/10.1016/j.foodchem.2015.04.017
• Alikord, M., Momtaz, H., Keramat, J., Kadivar, M., & Rad, A. H. (2018). Species identification and animal authentication in meat products: A review. Food Measure, 12(1): 145-155. https://doi.org/10.1007/s11694-017-9625-z.
• Bottero, M. T., Civera, T., Nucera, D., Rosati, S., Sacchi, P., & Turi, R. M. (2003). A multiplex polymerase chain reaction for the identification of cows’, goats’ and sheep’s milk in dairy products. Int Dairy J, 13(4), 277–282. https://doi.org/10.1016/S0958-6946(02)00170-X
• Chen, R. K., Chang, L. W., Chung, Y. Y., Lee, M. H., & Ling, Y. C. (2004). Quantification of cow milk adulteration in goat milk using high-performance liquid chromatography with electrospray ionization mass spectrometry Rapid Commun Mass Spectrom, 18(10), 1167-1171. https://doi.org/10.1002/rcm.1460
• Cheng, Y. H., Chen, S. D., & Weng, C. F. (2006). Investigation of Goats’ Milk Adulteration with Cows’ Milk by PCR. Asian-Aust J Anim, 19(10), 1503-1507. https://doi.org/10.5713/ajas.2006.1503
• Cosenza, G., Iannaccone, M., Gallo, D., & Pauciullo, A. (2019). A fast and reliable polymerase chain reaction method based on short interspersed nuclear elements detection for the discrimination of buffalo, cattle, goat, and sheep species in dairy products. Asian Austral J Anim, 32(6), 891-895. https://doi.org/10.5713/ajas.18.0459
• Deri̇nöz, A. N., Çufaoğlu, G., & Ayaz, N. D. (2021). Et türü tayininde kullanılan yöntemler. Akademik Et ve Süt Kurumu Dergisi, 1, 8–18.
• Di Pinto, A., Terio, V., Marchetti, P., Bottaro, M., Mottola, A., Bozzo, G., Bonerba, E., Ceci, E., & Tantillo, G. (2017). DNA-based approach for species identification of goat-milk products. Food Chemistry, 229, 93–97. https://doi.org/10.1016/j.foodchem.2017.02.067
• Drummond, M. G., Brasil, B. S. A. F., Dalsecco, L. S., Brasil, R. S. A. F., Teixeira, L. V., & Oliveira, D. A. A. (2013). A versatile real-time PCR method to quantify bovine contamination in buffalo products. Food Control, 29(1), 131-137. https://doi.org/10.1016/j.foodcont.2012.05.051
• Duarte-Vázquez, M. A., García-Ugalde, C. R., Álvarez, B. E., Villegas, L. M., García-Almendárez, B. E., Rosado, J. L., & Regalado, C. (2018). Use of urea-polyacrylamide electrophoresis for discrimination of A1 and A2 beta casein variants in raw cow’s milk. J Food Sci Technol, 55(5), 1942-1947. https://doi.org/10.1007/s13197-018-3088-z
• Food and Drug Administration (FDA) (1995). Center for food safety and applied nutrition. In Defect action level handbook. Washington Printing Office, Washington, DC.
• Golinelli, L. P., Carvalho, A. C., Casaes, R. S., Lopes, C. S. C., Deliza, R., Paschoalin, V. M. F., & Silva, J. T. (2014). Sensory analysis and species-specific PCR detect bovine milk adulteration of frescal (fresh) goat cheese. J Dairy Sci, 97(11), 6693–6699. https://doi.org/10.3168/jds.2014-7990
• González-Martínez, M. Á., Puchades, R., & Maquieira, Á. (2018b). Chapter 15-Immunoanalytical Technique: Enzyme-Linked Immunosorbent Assay (ELISA). In D. W. Sun (Ed.), Modern Techniques for Food Authentication (Second Edition) (pp. 617-657). Academic Press. https://doi.org/10.1016/B978-0-12-814264-6.00015-3
• Kara, R., & Demi̇rel, Y. N. (2016). Afyon kaymağı üretiminde kullanılan süt türünün Real-Time PCR ile belirlenmesi. Atatürk University Journal of Veterinary Sciences, 11(2), 185-190. https://doi.org/10.17094/avbd.77186
• Khatun, M. A, Hossain, A., Hossain, Md. S., Munshi, M. K., & Huque, R. (2021). Detection of species adulteration in meat products and Mozzarella-type cheeses using duplex PCR of mitochondrial cyt b gene: A food safety concern in Bangladesh. Food Chemistry: Molecular Sciences, 2, 100017. https://doi.org/10.1016/j.fochms.2021.100017
• Kumari, R., Rank, D. N., Kumar, S., Joshi, C. G., & Lal, S. V. (2015). Real time PCR an approach to detect meat adulteration. Buffalo Bull, 34(1), 124-129.
• López-Calleja, I. M., González, I., Fajardo, V., Hernández, P. E., García, T., & Martín, R. (2007). Application of an indirect ELISA and a PCR technique for detection of cows’ milk in sheep’s and goats’ milk cheeses. Int Dairy J, 17(1), 87–93. https://doi.org/10.1016/j.idairyj.2006.01.006
• López-Calleja, I., González, I., Fajardo, V., Rodríguez, M. A., Hernández, P. E., García, T., & Martín, R. (2004). Rapid detection of cows’ milk in sheeps’ and goats’ milk by a species-specific polymerase chain reaction technique. J Dairy Sci, 87(9), 2839-2845. https://doi.org/10.3168/jds.S0022-0302(04)73412-8
• Ma, A., Wang, Y., Liu, X. L., Zhang, H. M., Eamsobhana, P., Yong, H. S., & Gan, X. X. (2019). A filtration-based rapid test using a partially purified third-stage larval antigen to detect specific antibodies for the diagnosis of gnathostomiasis. Journal Helminthol, 93(1), 26–32. https://doi.org/10.1017/S0022149X17001080
• Mašková, E., & Paulíčková, I. (2006). PCR-based detection of cow’s milk in goat and sheep cheeses marketed in the Czech Republic. . Czech J Food Sci, 24(3), 127.
• Miller, G. D., Jarvis, J. K., McBean, L. D. (2006). Handbook of Dairy Foods and Nutrition (3nd ed.), CRC Press LLC, USA.
• Natonek-Wiśniewska, M., & Krzyścin, P. (2019). Detection of the species composition of food using mitochondrial DNA: challenges and possibilities of a modern laboratory. In Biochemical Analysis Tools-Methods for Bio-Molecules Studies. IntechOpen.
• Paszczyk, B., & Łuczyńska, J. (2020). The comparison of fatty acid composition and lipid quality indices in hard cow, sheep, and goat cheeses. Foods, 9(11), 1667. https://doi.org/10.3390/foods9111667
• Rancé, F., Grandmottet, X., & Grandjean, H. (2005). Prevalence and main characteristics of schoolchildren diagnosed withfood allergies in France. Clin Exp Allergy, 35(2), 167-172. https://doi.org/10.1111/j.1365-2222.2005.02162.x
• Rodrigues, N. P. A., Givisiez, P. E. N., Queiroga, R. C. R. E., Azevedo, P. S., Gebreyes, W. A., & Oliveira, C. J. B. (2012). Milk adulteration: Detection of bovine milk in bulk goat milk produced by smallholders in northeastern Brazil by a duplex PCR assay. J Dairy Sci, 95(5), 2749–2752. https://doi.org/10.3168/jds.2011-5235
• Rodríguez-Ramírez, R., González-Córdova, A. F., & Vallejo-Cordoba, B. (2011). Review: Authentication and traceability of foods from animal origin by polymerase chain reaction-based capillary electrophoresis. Analytica Chimica Acta, 685(2), 120-126. https://doi.org/10.1016/j.aca.2010.11.021
• Stănciuc (Sava), N., & Râpeanu, G. (2010). Identification of adulterated sheep and goat cheeses marketed in Romania by immunocromatographic assay. Food Agr Immunol, 21(2), 157–164. https://doi.org/10.1080/09540100903508683
• Ten-Doménech, I., Beltrán-Iturat, E., Herrero-Martínez, J. M., Sancho-Llopis, J. V., & Simó-Alfonso, E. F. (2015). Triacylglycerol analysis in human milk and other mammalian species: small-scale sample preparation, characterization, and statistical classification using HPLC-ELSD profiles. J Agr Food Chem, 63(24), 5761-5770. https://doi.org/10.1021/acs.jafc.5b01158
• Turkish Food Codex (TFC) (2017). Food labeling and consumer information regulation. Official Newspaper, 26 January 2017, 29960. Retrieved from https://www.resmigazete.gov.tr/eskiler/2017/01/20170126M1-6.htm; Accessed: December 15, 2021.
• Turkish Food Codex (TFC) (2019). Turkish Food Codex Communiqué on Drinking Milk. Official Newspaper, 27 February 2019, No: 2019/12, 2019. Retrieved from https://www.resmigazete.gov.tr/eskiler/2019/02/20190227-5.htm; Accessed: December 15, 2021.