سنجش مولکولی تقلب گوشت مرغ در همبرگر‌ممتاز به روش Simplex PCR و Duplex PCR
محورهای موضوعی :
علوم و صنایع غذایی
معصومه هاشم‌زادگان
1
,
سید ابراهیم حسینی
2
,
فرزانه تفویضی
3
,
منصور بیات
4
1 - دانش‌آموخته کارشناسی‌ارشد صنایع‌غذایی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران
2 - دانشیار گروه علوم و صنایع‌غذایی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران
3 - دانشیار گروه زیست‌شناسی، واحد پرند، دانشگاه آزاد اسلامی، پرند، ایران
4 - دانشیار گروه میکروبیولوژی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران
تاریخ دریافت : 1395/12/10
تاریخ پذیرش : 1396/12/20
تاریخ انتشار : 1397/04/01
کلید واژه:
گوشت مرغ,
همبرگر ممتاز,
Simplex-PCR,
Duplex PCR,
چکیده مقاله :
گوشت یک منبع پروتئین عالی و مغذی محسوب می شود اما به علت قیمت بالای این پروتئین حیوانی و دست‌یابی به سود اقتصادی بیشتر توسط برخی تولیدکنندگان، اغلب در فرآورده های گوشتی، تقلباتی نظیر استفاده از گونه های گوشتی با ارزش تجاری کمتر، مورد توجه است. در این تحقیق سعی شده تا پروتئین ارزان قیمتی نظیر گوشت مرغ به‌روش مولکولی شناسایی گردد. از 10 نوع همبرگر ممتاز صنعتی با درج عنوان تهیه شده از گوشت گاو و نمونه‌های شاهد، شامل گوشت خام گاو و مرغ،استخراج DNAانجام گرفت و سپس واکنش‌های Simplex PCR و Duplex PCR بر روی DNAهای حصول یافته با استفاده از پرایمرهای اختصاصی ویژه ژن سیتوکروم b گاو و s rRNA12 مرغ انجام شد. ژن‌های ویژه سیتوکروم b گاو و srRNA 12 مرغ به ترتیب قطعات bp 274 و bp 183 را طی واکنش‌های Simplex-PCR و Duplex-PCR در تمام نمونه های همبرگر ممتاز تکثیر کردند. تکثیر قطعه bp183 ویژه ژن مرغ طی واکنش‌های Simplex-PCR و Duplex-PCR، حاکی از جایگزینی تقلب مرغ با بخشی از گوشت گاو که ماده اصلی تمامی همبرگرهای ممتاز مورد آزمون است، می باشد.
چکیده انگلیسی:
Meat is considered an excellent and nutritious source of protein. However, due to the high cost of this animal protein and economic profit access of some producers, frauds like usage meat species of commercially lower value are noticeable. The aim of this study is to identify low-cost protein frauds such as chicken meat by the molecular method. DNA was extracted from 10 kinds of industrial premium beef burgers, and control samples involved raw beef and chicken meat. Then, simplex-PCR and duplex-PCR were run on the extracted DNA using specific primers of cytochrome b gene in beef and 12s rRNA gene in chicken. Specific genes of beef cytochrome b and chicken 12s rRNA amplified 274 bp and 183 bp fragments respectively in all premium burger samples in simplex-PCR and duplex-PCR. Amplification of fragments of 183 bp by simplex and duplex-PCR suggests adulteration of premium hamburgers by partial replacement of beef with chicken.
منابع و مأخذ:
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· Sakaridis, I., Ganopoulos, I., Argiriou, A. and Tsaftaris, A. (2013). A fast and accurate method for controlling the correct labeling of products containing buffalo meat using High Resolution Melting (HRM) analysis. Meat Science, 94: 84-88.
· Sambrook, J., Fritch, E. F., Maniatis, T. (1989). Molecular cloning. Cold spring Habor Laboratory Press, New York, pp. 3-15.
· Stamoulis, P., Stamatis, C., Sarafidou, T. and Mamuris, Z. (2010). Development and application of molecular markers for poultry meat identification in food chain. Food Control, 21: 1061-1065.
· Zilio Dinon, A., Treml, D., Souza de Mello. C. and Carolina Maisonnave Arisi, A. (2010). Monitoring of GMO in Brazilian processed meat and soy-based products from 2007 to 2008. Food Composition and Analysis, 23: 226 -229.
_||_ · Abbasi Fasarani, M., Hoseini, H., jahedkhaniki, Gh., Adibmoradi, M. Eskandari Gharabloo, S. (2012). The study of presence of illegal structure in industrial hamburgers contains 30 and 60 percent of meat by histological method and it̓s relationship with chemical indicators belonging to the connective tissue of meat. Journal of Nutrition and Food Technology, 7(5):311-318. [In Persian]
· Abd El-Nasser, M., Labieb, H. Y. and Abd El-Aziz, D. M. (2010). Detection of native and modified soybean in some meat products in Assiut city, Egypt. Assiut University Bull Environ Research, 13(1): 27-35.
· Amaral, J. S., Santos, C. G., Melo, V. S., Oliveira, M. B. P. P. and Mafra, I. (2014). Authentication of a traditional game meat sausage (Alheira) by species-specific PCR assays to detect hare, rabbit, red deer, pork and cow meats. Food Research International, 60: 140-145.
· Cammà, C., Di Domenico, M. and Monaco, F. (2012). Development and validation of fast Real-Time PCR assays for species identification in raw and cooked meat mixtures. Food Control, 23: 400-404.
· Chun Chi, L., Lai Ling, F., Po Kwok, Ch., CheukMana, L., Kwok Fai, Ch. and Shuk Han, Ch. (2014). A rapid low-cost high-density DNA-based multi-detection test for routine inspection of meat species. Meat Science, 96: 922–929.
Dalmasso, A., Fontanellab, E., Piattib, P., Civeraa, T., Rosatic, S. and Bottero, M. T. (2004). A multiplex PCR assay for the identification of animal species in feedstuffs. Molecular and Cellular Probes, 18: 81–87.
· Doosti, A., Ghasemi Dehkordi, P. and Rahimi, E. (2014). Molecular assay to fraud identification of meat products. Food Science Technology, 51(1): 148-152.
· Eaqub Ali, M., Hashim, U., Mustafa, S. and Man, Y. (2012). Swine-specific PCR-RFLP assay targeting mitochondrial cytochrome b gene for semi quantitative detection of pork in commercial meat products. Food Analysis Methods, 5: 613–623.
· Flores-Munguia, M. E., Bermudez-Almada, M. C. and Vazquez-Moreno, L. (2000). A research note: Detection of adulteration in processed traditional meat products. Muscle Foods, 11: 319-325.
· Forte, V. T., DiPinto, A., Martino, C., Tantillo, G. M., Grasso, G. and Schena, F. P. (2005). A general multiplex-PCR assay for the general detection of genetically modified soya and maize. Food Control, 16: 535–539.
· Gazalli, H., Malik, A. H., Jalal, H., Afshan, S., Mir, A. and Ashraf, H. (2013). Methods of identification of meat species. Food Nutrition And Safety, 3(2): 90-110.
· Ghovvati, S., Nassiri, M. R., Mirhoseini, S. Z., Heravi Moussavi, A. and Javadmanesh, A. (2009). Fraud identification in industrial meat products by multiplex PCR assay. Food Control, 20: 696–699.
· Hoseini, H., Barazandegan, Kh., Akhoondzadeh, A., Shemshadi, B., tavakoli, H. and Khaksar, R. (2009). Determination the kind of meat content of Patties marketed in Tehran in 2009.Journal of Food Science and Technology, 3: 95-100. [In Persian]
· Institute of Standards and Industrial Research of Iran. (1991). Frozen raw hamburger properties. 3th revision, ISIRI No. 2304. [In Persian]
· Karabasanavar, S. N., Singh, S. P., Kumar, D. and Shebannavar, S. (2014). Detection of pork adulteration by highly-specific PCR assay of mitochondrial D-loop. Food Chemistry, 145: 530–534.
· Kim, J., Jeong, D., Kim, Y., Kwon, Y., Rhee, G., Zhang, D. et al., (2013). Development of a multiplex PCR method for testing six GM soybean events. Food Control, 31: 366-371.
· Kesmen, Z., Celebi, Y., Güllüce, A. and Yetim, H. (2013). Detection of seagull meat in meat mixtures using real-time PCR analysis. Food Control, 34: 47-49.
· Mafra, I., Ferreira, I. and Oliveira, B. (2008). Food authentication by PCR-based methods, Springer-Verlag. Europian Food Research and Technology, 227: 649–665.
· Mane, B. G., Mendiratta, S. K. and Tiwari, A. K. (2009). Polymerase chain reaction assay for identification of chicken in meat and meat products. Food Chemistry, 116: 806–810.
· Matsunaga, T., Chikuni, K., Tanabeb, R., Muroyab, S., Shibata, K., Yamadaa, J. et al., (1999). A quick and simple method for the identication of meat species and meat products by PCR assay. Meat science, 51: 143-148.
· Nollet, L. (2007). Handbook of Meat, Poultry and Seafood Quality, Blackwell, Ames, pp. 587-694.
· Pelt-Verkuil, E. V., Belkum, A. V. and P. Hays, J. (2008). Principles and Technical Aspects of PCR Amplification. Trends Food Science and Technology, 9: 380–388.
· Safdar, M. and Abasıyanık, M. F. (2013). Development of fast multiplex real-time PCR assays based on EvaGreen fluorescence dye for identification of beef and soybean origins in processed sausages. Food Research International, 54: 1652–1656.
· Sakaridis, I., Ganopoulos, I., Argiriou, A. and Tsaftaris, A. (2013). A fast and accurate method for controlling the correct labeling of products containing buffalo meat using High Resolution Melting (HRM) analysis. Meat Science, 94: 84-88.
· Sambrook, J., Fritch, E. F., Maniatis, T. (1989). Molecular cloning. Cold spring Habor Laboratory Press, New York, pp. 3-15.
· Stamoulis, P., Stamatis, C., Sarafidou, T. and Mamuris, Z. (2010). Development and application of molecular markers for poultry meat identification in food chain. Food Control, 21: 1061-1065.
· Zilio Dinon, A., Treml, D., Souza de Mello. C. and Carolina Maisonnave Arisi, A. (2010). Monitoring of GMO in Brazilian processed meat and soy-based products from 2007 to 2008. Food Composition and Analysis, 23: 226 -229.
