Quantitative detection of chicken meat routine mislabeling in emulsion type sausages and burgers by SYBR green real time PCR assay

Document Type: Full Article

Authors

1 Department of Food Hygiene and Public Health, School of Veterinary Medicine, Shiraz University, Shiraz, I. R. Iran

2 Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, I. R. Iran

Abstract

ABSTRACT- Today, the authenticity of meat products with less costly and desirable species has increased. Therefore and considering religious, economicalor public health concerns, proper actions should be taken to prevent such frauds. In this study, real time PCR assay was applied for rapid, sensitive and specific identification and quantification of chicken tissue in meat products. Specific primers were designed on the 16SrRNA chicken mitochondrial genes. The conventional PCR and SYBR Green RT-PCR were employed on DNA extracted from 150 samples, sausages and burgers. The results showed the presence of misused chicken meat in the sausages and burgers were 84% and 26%, respectively. Additionally, the Q-PCR assay was able to detect estimate rates of 10% to 50% of chicken meat in the products. The detection limit of the method could be quantitatively used by regulation authorities to control the quality of meat products.

Keywords


Article Title [Persian]

شناسایی و اندازه‌گیری مقادیر غیر مجاز خمیر مرغ در محصولات گوشتی شامل سوسیس و برگر با استفاده از روش سایبرگرین

Authors [Persian]

  • لیلا لک‌زاده 1
  • سید شهرام شکرفروش 1
  • مهدی فاضلی 2
1 دانشگاه شیراز
2 دانشگاه شیراز
Abstract [Persian]

چکیده- امروزه توجه به تولید محصولات گوشتی بدون تقلب و با هزینه کمتر افزایش یافته است. بنابراین با در نظر گرفتن نگرانی‌های مربوط به سلامت عمومی اعتقادات مذهبی و مسائل اقتصادی، لازم است اقدامات مناسب جهت ممانعت از این تقلبات به عمل آید. در مطالعه حاضر با استفاده از آزمایش Real Time PCR به عنوان یک روش سریع، حساس و اختصاصی، به شناسایی و اندازه‌گیری میزان بافت‌های مرغی در محصولات گوشتی با برچسب تهیه شده از گوشت قرمز پرداخته شده است. بدین منظور، بر اساس ژن 16 rRNA ، پرایمرهای اختصاصی آماده گردید و با روشهای PCR و RT-PCR بر روی تعداد 150 DNA استخراج شده از نمونه‌های سوسیس و برگر انجام گردید. نتایج حاصل از مطالعه حاضر نشان دهنده وجود 84% و 26% بافت مرغ به ترتیب در نمونه‌های سوسیس و برگر می‌باشد. به علاوه روش RT-PCR(Q-PCR) قادر به تشخیص تقریبی 5-10% بافت مرغ در این محصولات است. این محدوده تشخیص توسط روش فوق می‌تواند به طور موثری توسط مراجع ذی‌صلاح به منظور کنترل کیفیت محصولات گوشتی مورد استفاده قرار گیرد.

Keywords [Persian]

  • واژه های کلیدی:
  • مرغ
  • سایبرگرینRT-PCR
  • 16S rRNA
  • برگر
  • سوسیس
Aguado, V., Vitas, A.I., & Garcia Jalon, I. (2001). Random amplified polymorphic DNA typing applied to the study of cross-contamination by Listeria monocytogenesin processed food products. Journal of Food Protection, 64 (5),716–720.

Ballin, N.Z. (2010). Authentication of meat and meat products. Meat Science, 86, 577–587.

Ballin, N.Z., Vogensen, F.K., & Karlsson, A.H. (2009). Species determination – Can we detect and quantify meat adulteration? Meat Science, 83, 165–174.

Brodmann, P.D., & Moor, D. (2003). Sensitive and semi-quantitative Taq Man real-time polymerase chain reaction systems for the detection of beef (Bostaurus) and the detection of the family mammalian in food and feed. Meat Science, 65, 599–607.

Calvo, J.H., Zaragoza, P., & Osta, R. (2001). A quick and more sensitive method to identify pork in processed and unprocessed food by PCR amplification of a new specific DNA fragment. Journal of Animal Science, 79, 2108-2112. 

Fajardo, V., Gonzalez, I. Martın, R. Rojas, M, Hernández, P.E., & García, T.  (2008). Real-time PCR for detection and quantification of red deer (Cervus elaphus), fallow deer (Damadama), and roe deer (Capreolu scapreolus) in meat mixtures. Meat Science, 79, 289–298.

Fajardo, V., Gonzalez, I., Rojas, M., Garcıa, T., & Martın, R. (2010). A review of current PCR-based methodologies for the authentication of meats from game animal species. Trendsin Food Science and Techology, 21, 408-421.

Grish, P.S., Anjaneyulu, A.S.R., Viswas, K.N., Shivakumar, B.M., Anand, M., Patel, M., & Sharma, B. (2005). Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene. Meat Science, 70, 107-112.

Haunshi, S., Basumatary, R., Grish, P.S., Doley, S., Bardoloi, R.K., & Kumar, K. (2009). Identification of chicken, duck, pigeon and pig meat by species-specific markers of mitochondrial origin. Meat Science, 83, 454-459.

Hopewood, A.J., Fairbrother, K.S., Lockley, A.K., & Bardsley, R.G. (1999).  An actiongene-related polymerase chain reaction (PCR) test for identification of chicken in meat mixtures. Meat Science, 53, 227-231.

Kesmen, Z. (2005). Research on the meat species identification in fresh and processed meat products by using PCR techniques.Ph.D. Dissertation, Ataturk University, Erzurum.

Kesmen, Z., Gulluce, A., Sahin, F., & Yetim, H. (2009).  Identification of meat species by TaqMan- based real-time PCR assay. Meat Science, 82, 444- 449.

Kesmen, Z., Sahin, F., & Yetim, H. (2007). PCR assay for the identification of animal species in cooked sausages. Meat Science, 77, 649-653.

Kesmen, Z., Yetim, H., & Sahin, F. (2006). PCR assay for the identification of animal species in Turkish fermented sausages. In 1stEgyptian–Jordanian conference on biotechnology and sustainable development:Current status and future scenarios,11–14 December, (pp. 151–155), Cairo, Egypt.

López Andreo, M., Lugo, L., Garrido Pertierra, A., Prieto, M.I., & Puyet, A. (2005). Identification and quantitation of species in complex DNA mixtures by real-timepolymerase chain reaction. Analytical Biochemistry, 339, 73-82.

Martin, I., Garcia, T., Fajardo, V., Rojas, M., Pegels, N., & Hernandez, P.H. (2009). SYBR-Green real-time PCR approach for the detection and quantification of pig DNA in feed stuffs. Meat Science, 82, 252-259.

Matsonaga, 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.

Rodrıguez, M.A., Garcıa, T., Gonzalez, I., Hernandez, P., & Martın, R.  (2005). TaqMan real-time PCR for the detection and quantitation of pork in meat mixtures. Meat Science, 70, 113–120.

Rojas, M., González, I., Pavón, M.A., Pegels, N., Hernández, P., García, T., & Martin, R. (2011). Application of a real-time PCR assay for the detection of ostrich (Struthio camelus) mislabelling in meat products from the retail market. Food Control, 22, 523-531.

Rojas, M., González, I., Fajardo, V., Martín, I., Hernández, P., García, T., & Martín, R. (2009). Authentication of meats from quail (Coturnixcoturnix), pheasant (Phasianus colchicus), partridge (Alectoris spp.), and guinea fowl (Numidam eleagris) using polymerase chain reaction targeting specific sequences from the mitochondrial 12S rRNA gene. Food Control, 20, 896-902.

Sawyer, J., Wood, C., Shanahan, D., Gout, S., & McDowell, D. (2003). Real-time PCR quantitative meat species testing.  Food Control, 14, 579–583.

Stamoulis, P., Stamatis, C., Sarafidou, T., & Mamuris, Z. (2010). Development and application of molecular markers for poultry meat identification in food chain. Food Control, 21, 1061-1065.

Verkaar, E.L.C., Nijman, I.J., Boutaga, K., & Lenstra, J.A. (2002). Differentiation of cattle species in beef by PCR-RFLP of mitochondrial and satellite DNA. Meat Science, 60, 365–369.

Wang, R.F., Myers, M.J., Campbell, W., Cao, W.W., Paine, D., & Cerniglia, C.E. (2000). A rapid method for PCR detection of bovine materials in animal feedstuffs. Molecular and Cellular Probes, 14 (1), 1-5.