Real-Time PCR Tekniği ile Çeşitli Et Ürünlerinde Tavuk ve Sığır Eti Oranlarının Kantitatif Tyini

Öz

Et ürünleri üretiminde kullanılan et türlerinin tür bazında ve oransal olarak doğru belirlenmesi tüketicinin aldatılmasının önlenmesi, halk sağlığının korunması ve üreticiler arası haksız rekabetin önlenmesi açısından büyük önem taşımaktadır.Yapılan bu çalışmada, hem tür analizi hem de kantitatif analiz yapabildiği üretici firma tarafından ifade edilen SureFood Animal QUANT Beef ve SureFood Animal QUANT Chicken real- time PCR kitleri kullanılmştır. Çalışmada deneysel olarak belirli oranlarda (%10 sığır eti + %90 tavuk eti, %20 sığır eti + %80 tavuk eti, %30 sığır eti + %70 tavuk eti, %50 sığır eti + %50 tavuk eti, %20 tavuk eti + %80 sığır eti, %30 tavuk eti + %70 sığır eti olmak üzere) sığır eti + tavuk eti karışımlarının yanı sıra farklı oranlarda sığır ve tavuk etinden hazırlanmış et ürünleri (salam, sosis ve sucuk) test materyali olarak kullanılmıştır. Et karışımlarından ve et ürünlerinden DNA ekstraksiyon işlemleri SureFood Prep Animal DNA ekstraksiyon kiti (Congen S 1003, Berlin, Almanya) ile yapılmıştır. DNA ekstraksiyonunu takiben Real-Time PCR tekniği ile et oranları tayin edilmeye çalışılmıştır. PCR analizleri Light Cycler-2 (Roche) cihazında gerçekleştirilmiştir. Çalışma sonuçlarına göre, kullanılan SureFood Animal QUANT Beef and SureFood Animal QUANT Chicken real-time PCR kitlerinin deneysel olarak farklı oranlarda sığır ve tavuk etinden hazırlanmış et karışımları ile yine çeşitli sığır ve tavuk eti karışımlarından yapılan salam, sosis ve sucuk örneklerinde sığır ve tavuk eti oranlarını kantitatif olarak doğru tespit edemediği ortaya konulmuştur.

Anahtar Kelimeler

• Etlik Veteriner Mikrobiyoloji Dergisi
• Et Ürünleri
• Et Türleri
• Kantitatif Analiz
• Real-Time PCR
• Yıldız AYAZ
• Naim Deniz AYAZ
• Mihriban AKSOY
• Yusuf Ziya KAPLAN
• 2013

Quantitative Detection of Chicken and Beef Meat From Meat Products by Real-Time PCR

Öz

Quantitative detection of meat species that are used in production of meat products has a great importance for preventing consumers from adulteration, protection of public health and prevention of unfair competition between producers. In this study, SureFood Animal QUANT Beef and SureFood Animal QUANT Chicken real-time PCR kits were used to detect meat species quantitatively. For this purpose, various proportions (10% bovine + 90% chicken, 20% bovine + 80% chicken, 30% bovine + 70% chicken, 50% bovine + 50% chicken, 80% bovine + 20% chicken, 70% bovine + 30% chicken) of bovine and chicken meat mixtures and meat products (salami, sausage and sucuk from different bovine + chicken meat) were experimentally prepared. After DNA extraction process of these meat mixtures and products, Real-Time PCR technique was used for the quantitative detection of meat species. SureFood Prep Animal DNA extraction kit (Congen S 1003, Berlin, Germany) was used for the extraction of DNA from mixtures. PCR analyses were carried out with Light Cycler-2 (Roche, Germany). According to the real-time PCR results, SureFood Animal QUANT Beef and SureFood Animal QUANT Chicken kits were not able to detect the rates of meat species in meat mixtures (beef and chicken) and meat products (salami, sausage and sucuk) accurately.

Kaynakça

1. Ayaz Y, Ayaz ND, Erol I, (2006). Detection of species in meat and meat products using enzyme-linked immunosorbent assay. J Muscle Foods. 17, 214-220.
2. Berger RG, Mageau RP, Schwab B, Johnston RW, (1988). Detection of poultry and pork in cooked and canned meat foods by enzyme linked immunosorbent assays. J AOAC Int. 71, 406-409.
3. Chikuni K, Tabata T, Kosugiyama M, Monma M, (1994). Polymerase chain reaction assay for detection of sheep and goat meats. Meat Scie. 37, 337-345.
4. Chisholm J, Conyers C, Booth C, Lawley W, Hird H, (2005). The detection of horse and donkey using real-time PCR. Meat Scie. 70, 727-732.
5. Chung ER, Kim YS, Han SK, (2000). Identification of Hawoon meat using PCR-RFLP marker of MCIR gene associated with bovine coat color. J Animal Sci Technol. 42, 379-380.
6. Dooley JJ, Paine KE, Garrett SD, Brown HM, (2004). Detection of meat species using TaqMan real-time PCR assays. Meat Scie. 68, 431-438.
7. Ganai TAS, Sing RK, Butchaiah G. (2000). DNA amplification fingerprinting of cattle and buffalo genome by RAPD-PCR utilizing arbitrary oligonucleotide primers. Buffalo J. 3, 331-339.
8. Girish PS, Anjaneyulu ASR, Viswas KN, Anand M, Rajkumar N, Shivakumar BM, Et Al., (2004). Sequence analysis of mitochondrial 12S rRNA gene can identify meat species. Meat Sci. 66, 551-556.

9. Guoli Z, Mingguang Z, Zhiang Z, Hongsheng O, Qiang L, (1999). Establishment and application of a polymerase chain reaction for the identification of beef. Meat Sci. 51, 233-236.
10. Günşen U, Aydın A, Ovalı BB, Coşkun Y, (2006). Detection of Different Meat Species in Raw Meat and Cooked Meat Products Using ELISA Technique. J Vet Fac Vet Med. Istanbul Univ. 32, 45-52.
11. Hsieh YHP, Chen FC, Sheu SC, (1997). A. AES research developing simple, inexpensive tests for meat products., Highlights of Agricultural Research, 44(2), Summer.
12. Hsieh YHP, Johnson MA, Wetzstein CJ, Green NR, (1996). Detection of species adulteration in pork products using agar gel immunodiffusion and enzyme linked immunosobent assay. J Food Quality. 19, 1-9.
13. Hsieh YHP, Woodward BB, Ho SH, (1995). Detection of species substitution in raw and cooked meats using immunoassays. J Food Quality. 58, 555-559.
14. Köppel R, Ruf J, Rentsch J, (2011). Multiplex real-time PCR for the detection and quantification of DNA from beef, pork, horse and sheep. Eur Food Res Technol. 232, 151-155.
15. Kremar P, Rencova E, (2005). Quantitative Detection of Species-Specific DNA in Feedstuffs and Fish Meal. Journal of Food Protec. 68, 1217-1221.
16. Lahiff S, Glennon M, Lyng J, Smith T, et al. (2001). Real- time Polymerase chain reaction detection of bovine DNA in meat and bone meal samples. J Agric Food Chem. 7, 1158-1165.
17. Lombardo F, Galli A, Bongioni G, (2004). Real-Time PCR Approach for Detection of Bovine DNA in Animal Feedstuffs, Atti del 39 Simposio Internazionale di Zootecnica “Meat scie and research”, 10 giugno Roma.
18. Matsunga T, Chikuni K, Tanabe R, Muroya H, Shibata K, Yamada J, Et Al. (1998). Determination of mitochondrial cytocrome b gene sequence for red deer (Cercus elaphus) and the differentiation of closely related deer meats. Meat Sci. 49, 379-385.
19. Necidova L, Recencova E, Svoboda L, (2002). Counter immunoelectrophoresis: a simple method for detection of species specific muscle proteins in heat processed products. Vet Med Czech. 47, 143-147.
20. Reddy PM, Reddy VSL, Rao ZS, Murthy GK, (2000). Identification of origin of fresh,cooked, and decomposed meats by using brain antigens. J Food Sci. Technol. 37, 201-203.
21. Rencova F, Necidova L, Svoboda L, (2000). Identification by ELISA of poultry, horse, kangaroo, and rat muscle specific proteins in heat processed products. Vet Med Czech. 45, 353-356.
22. Shericar AT, Karkare UD, Khot JB, Jayarao BM, Bhilegaonkar KN, (1993). Studies on termostable antigens, production of species specific antiadrenal sera and comparison of immunological techniques in meat speciation. Meat Sci. 33, 121-136.
23. Soares S, Amaral JS, Mafra I, Oliveira MBPP, (2010). Quantitative detection of poultry meat adulteration with pork by a duplex PCR assay. Meat Sci. 85, 531-536.
24. Tremlova B, (2000). Histologischer Nachweis von Knochenpartikein in Fleischprodukten. Fleiswirtsch. 80, 73-74.