Authentication of Meat Species in Sucuk by Multiplex PCR

Year 2015, Volume: 41 Issue: 1, 6 - 11, 05.01.2015

Osman İlhak , Hüsnü Güran

Abstract

The identification of meat species used in meat products is important by reason of economic considerations, religious factors, verification of label, and prevention of unfair-market competition. In this paper, multiplex PCR method was experienced for routine detection of equine (horse and donkey), poultry (chicken and turkey), pig and cattle meat in sucuk (sausage). The primers used for these animals generated specific fragments, and they did not show cross reactions with the DNA from the other genus of animal. After multiplex PCR was successfully optimized, a field study was carried out to investigate the presence of horse, donkey, chicken, turkey and pig meat in 50 sucuks (30 beef and 20 beef + poultry) collected from markets. The result of the field study indicated that 23.3% of 30 beef sucuk samples were containing poultry meat. None of the 50 sucuk samples was containing pig meat, but one (2%) of the samples generated equine fragment. The present study showed that the multiplex PCR method can be used for routine analysis of meat species identification, verification and control of label information of meat products.

Keywords

Multiplex PCR, species identification, sucuk

Multipleks PCR ile Sucuklarda Et Türlerinin Doğruluğunun Kanıtlanması

Abstract

Et ürünlerinde kullanılan et türlerinin saptanması ekonomik sebepler, dini faktörler, etiketin doğrulanması ve haksız rekabetin önlenmesi açısından önemlidir. Bu çalışmada, sucuklarda tek tırnaklı (at ve eşek), kanatlı (tavuk ve hindi), domuz ve sığır etinin rutin tespiti için multipleks PCR metodu denendi. Bu hayvanların DNA’larının tespitinde diğer hayvan türleri ile çapraz eşleşme göstermeyen türe özgü primerler kullanıldı. Multipleks PCR başarılı bir şekilde optimize edildikten sonra, marketlerden toplanmış 50 adet sucukta (30 adeti sığır eti, 20 adeti sığır eti-kanatlı eti karışımı) at, eşek, tavuk, hindi ve domuz etinin varlığının araştırıldığı bir saha çalışması gerçekleştirildi. Saha çalışmasının sonuçları sığır etinden üretilmiş 30 adet sucuğun %23,3’ünün kanatlı eti içerdiğini gösterdi. Elli adet sucuk örneğinin hiçbirinde domuz eti bulunmadı fakat bir örnekten (%2) tek tırnaklı DNA parçası çoğaltıldı. Bu çalışma multipleks PCR metodunun et ürünlerinde et türlerinin belirlenmesi, doğrulanması ve etiket bilgilerinin kontrolü için rutin olarak kullanılabileceğini gösterdi.

Keywords

Multipleks PCR, tür tayini, sucuk

References

• Ali, M.E., Hashim, U., Mustafa, S., Man, Y.B.C., 2012. Swine- Specific PCR-RFLP assay targeting Mitochondrial Cytochrome B gene for semiquantitative detection of pork in commercial meat products. Food Analytical Methods 5, 613-623.
• Anonymous, 2012. Turkish Food Codex Notification No. 2012/74. (Türk Gıda Kodeksi Et ve Et Ürünleri Tebliği (Tebliğ No: 2012/74).
• Ayaz, Y., Ayaz, N.D., Erol, İ., 2006. Detection of species in meat and meat products using Enzyme-linked immunosorbent assay. Journal of Muscle Foods 17, 214-220.
• Bai, W., Xu, W., Huang, K., Yuan, Y., Cao, S., Luo, Y., 2009. A novel common primer multiplex PCR (CP-M-PCR) method for the simultaneous detection of meat species. Food Control 20, 366-370.
• 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. International Dairy Journal 13, 277-282.
• Çetin, O., Bingöl, E.B., Akkaya, H., 2008. The microbiological, serological and parasitological quality of çig kofte (raw meatball) and its lettuce marketed in Istanbul. Polish Journal of Environmental Studies 17, 701-706.
• Çetin, O., Bingöl, E.B., Çolak, H., Ergün, O., Demir, C., 2010. The microbiological, serological and chemical quality of mincemeat marketed in İstanbul. Turkish Journal of Veterinary and Animal Sciences 34, 407-412.
• Dalmasso, A., Fontanella, E., Piatti, P., Civera, T., Rosati, S., Bottero, M., 2004. A multiplex PCR assay for the identification of animal species in feedstuffs. Molecular and Cellular Probes 18, 81-87.
• Ercoşkun, H., Özkal, S.G., 2011. Kinetics of traditional Turkish sausage quality aspects during fermentation. Food Control 22, 165-172.
• Fajardo, V., Gonzalez, I., Lopez-Calleja, I., Martin, I., Hernandez, P.E., Garcia, T., Martin, R., 2006. PCR– RFLP authentication of meats from red deer (Cervus elaphus), fallow deer (Dama dama), roe deer (Capreolus capreolus), cattle (Bos taurus), sheep (Ovis aries), and goat (Capra hircus). Journal of Agricultural and Food Chemistry 54, 1144-1150.
• Ghovvati, S., Nassiri, M.R., Mirhoseini, S.Z., Heravi Moussavi A., Javadmanesh, A., 2009. Fraud identification in industrial meat products by multiplex PCR assay. Food Control 20, 696-699.
• Günşen, U., Aydın, A., Ovalı, B.B., Coşkun, Y., 2006. Çiğ et ve ısıl işlem görmüş et ürünlerinde ELISA tekniği ile farklı et türlerinin tespiti. İstanbul Üniversitesi Veteriner Fakültesi Dergisi 32, 45-52.
• Haunshi, S., Basumatary, R., Girish, P.S., Doley, S., Bardoloj, R.K., Kumar, A., 2009. Identification of chicken, duck, pigeon and pig meat by species-specific markers of mitochondrial origin. Meat Science 83, 454-459.
• İlhak, O.İ., Arslan, A., 2007. Identification of meat species by polymerase chain reaction (PCR) technique. Turkish Journal of Veterinary and Animal Sciences 31, 159- 163.
• Kesmen, Z., Sahin, F., Yetim, H., 2007. PCR assay for the identification of animal species in cooked sausages. Meat Science 77, 649-653.
• Koh, M.C., Lim, C.H., Chua, S.B., Chew, S.T., Phang, S.T.W., 1998. Random amplified polymorphic DNA (RAPD) fingerprints for identification of red meat animal species. Meat Science 48, 275-285.
• Lahiff, S., Glennon, M., O’Brien, L., Lyng, J., Smith, T., Maher, M., Shilton, N., 2001. Species-specific PCR for the identification of ovine, porcine and chicken species in meat and bone meal (MBM). Molecular and Cellular Probes 15, 25-35.
• Mane, B.G., Mendiratta, S.K., Tiwari, A.K., Sharma, B.D., Bhilegaokar, K.N., Anjaneyulu, A.S.R., 2011. Detection of pork in admixed meat and meat products by species-specific PCR technique. The Indian Journal of Animal Sciences 81, 1178-1181.
• Matsunaga, T., Chikuni, K., Tanabe, R., Muroya, S., Shibata, K.,Yamada, J., Shinmura, Y., 1999. A quick and simple method for the identification of meat species and meat products by PCR assay. Meat Science 51, 143- 148.
• Murugaiah, C., Noor, Z.M., Mastakim, M., Bilung, L.M., Selamat, J., Radu, S., 2009. Meat species identification and halal authentication analysis using mitochondrial DNA. Meat Science 83, 57-61.
• Özpınar, H., Tezmen, G., Gökçe, İ., Tekiner, İ.H., 2013. Detection of animal species in some meat and meat products by comparatively using DNA microarray and real time PCR methods. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 19, 245-252.
• Pegels, N., Gonzalez, I., Martin, I., Rojas, M., Garcia, T., Martin, R., 2011. Applicability assessment of a real time PCR assay for the specific detection of bovine, ovine and caprine material in feedstuffs. Food Control 22, 1189-1196.
• Sakalar, E., Abasiyanik, M.F., 2012. The development of dublex real-time PCR based on SYBR green florescence for rapid identification of ruminant and poultry origins in foodstuff. Food Chemistry 130, 1050-1054.
• Şakalar, E., Abasiyanik, M.F., 2011. Qualitative analysis of meat and meat products by multiplex polymerase chain reaction (PCR) technique. African Journal of Biotechnology 10, 9379-9386.
• Ulca, P., Balta, H., Cagin, I., Senyuva, H.Z., 2013. Meat species identification and Halal authentication using PCR analysis of raw and cooked traditional Turkish food. Meat Science 94, 280-284.
• Türk, N., Kafa, B., İzan, Y., 2005. Et ve et ürünlerinde tür tayini. 5. Gıda Kongresi, 19-21 Nisan İzmir, Türkiye.
• Türkyılmaz, Ö., Kafa, B., İzan, Y., Sava, Ş., 2009. Çiğ et ve et ürünlerinde AGID yöntemi ile türlerin tespiti. Bornova Veteriner Kontrol Araştırma Enstitüsü Dergisi 31, 15- 20.
• Yalçın, H., Alkan, G., 2012. Investigation of the presence of horse and pig meat in meat and meat products using uhlenhuth precipitation ring, agar gel immuno- diffusion and enzyme-linked immuno sorbent assay techniques. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 18, 923-927.
• Zhang, C.L., Fowler, M.R., Scott, N.W., Lawson, G., 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.