* Corresponding Author
AUTHENTICATION OF RAWCHICKEN MEAT FROM PORKCONTAMINATION USING GENE CYT-BWITH DUPLEX-POLYMERASE CHAIN REACTION ANALYSIS
https://doi.org/10.21059/buletinpeternak.v41i2.13376 Bayu Setya Hertanto(1), Rizky Aulia Fitra(2), Lilik Retna Kartikasari(3), Muhammad Cahyadi(4*)
(1) Department of Animal Science Faculty of Agriculture Universitas Sebelas Maret
(2) Department of Animal Science Faculty of Agriculture Universitas Sebelas Maret
(3) Department of Animal Science Faculty of Agriculture Universitas Sebelas Maret
(4) Department of Animal Science Faculty of Agriculture Universitas Sebelas Maret
(*) Corresponding Author
Abstract
Halal is one of important aspects in consumer protection. Meat and processed meat products are food that should be controlled strictly because those are prone to be adulterated by pork contamination. Therefore, it is necessary to provide detection technique which is accurate, fast and cheap. The objective of this research was to identify the presence of impurities of pork meat on raw chicken meat using gene Cyt-b with duplex-PCR analysis. This research used six samples of raw chicken meat and raw pork. Raw chicken meat was bought from supermarkets in the city of Surakarta and raw pork was obtained from pig slaughterhouse. The percentage of raw pork contamination on raw chicken meat was designed as much as 1, 5, 10, and 25%, respectively. The DNA genome was isolated according to DNA isolation protocol from Genomic DNA Mini Kit. In addition, duplex-PCR was performed based on protocol of KAPA2G Fast Multiplex PCR kit. The data was descriptively analyzed by directly looking the DNA bands on the gel documentation apparatus. The result showed that specific DNA bands for chicken and pig were completely appeared on 1.5% of agarose gels. Duplex-PCR detect contamination of pork on raw meat of chicken at all contamination levels. This research proved that the duplex-PCR detect the contamination of pork until the level of 1%.
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Aida, A. A., Y. B. Che Man, C. M. V. L. Wong, A. R. Raha, and R. Son. 2005. Analysis of raw meats and fats of pigs using polymerase chain reaction for halal authentication. Meat Sci. 69: 47-52.
Ali, M. E., U. Hashim, S. Mustafa, Y. B. Che Man, T. S. Dhahi, M. Kashif, Md. Kamal Uddin, and S. B. Abd Hamid. 2012. Analysis of pork adulteration in commercial meatballs targeting porcine-specific mitochondrial cytochrome b gene by TaqMan probe real-time polymerase chain reaction. Meat Sci. 91: 454-459.
Ballin, N. Z. 2010. Authentication of meat and meat products. Meat Sci. 86: 577-587.
Bottero, M. T. and A. Dalmasso. 2011. Animal species identification in food products: Evolution of biomolecular methods. Vet. J. 190: 34-38.
Che Man, Y. B., A. A. Aida, A. R. Raha, and R. Son. 2007. Identification of pork derivatives in food products by species-specific polymerase chain reaction (PCR) for halal verification. Food Control 18: 885-889.
Dooley, J. J., K. E. Paine, S. D. Garrett, and H. M. Brown. 2004. Detection of meat species using TaqMan real-time PCR assays. Meat Sci. 68: 431-438.
Erwanto, Y., M. Z. Abidin, A. Rohman and Sismindari. 2011. PCR-RFLP using BseDi enzyme for pork authentication in sausage and nugget product. Media Peternakan 3: 14-18.
Faatih, M. 2009. Isolasi dan digesti DNA kromosom. Jurnal Penelitian Sains dan Teknologi 10: 61-67.
Girish, P. S., A. S. R. Anjaneyuhu, K. N. Viswas, B. M. Shivakumar, M. Anand, M. Patel, and B. Sharma. 2005. Meat spesies identification by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene. Meat Sci. 70: 107-112.
Jain, S., M. N. Brahmbhait, D. N. Rank, C. G. Joshi and J. V. Solank. 2007. Use a cytochrome b gene variability in detecting meat species by multiplex PCR assay. Indian J. Anim. Sci. 77: 880-881.
Kesmen, Z., A. Gulluce, F. Sahin, and H. Yetim. 2009. Identification of meat species by taqman based real time PCR assay. Meat Sci. 82: 444-449.
Koppel, R., J. Ruf, and J. Rentsch. 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.
Ling-Sun, Y. and C. Sheng Lin. 2003. Establishment and application of a fluorescent polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identifying porcine, caprine and bovine meats, chemistry. Agr. Food Chem. 51: 1771-1776.
Matsunaga, T., K. Chikuni, R. Tanabe, S. Muroya, K. Shibata, J. Yamada, and Y. Shinmura. 1999. A quick and simple method for the identification of meat species and meat products by PCR assay. Meat Sci. 51: 143-148.
Murugaiah, C., Z. M. Noor, M. Mastakim, L. M. Bilung, J. Selamat, and R. Son. 2009. Meat species identification and halal authentication analysis using mitochondrial DNA. Meat Sci. 83: 57-61.
Nakyinsige, K., Y. B. Man, and A. Q. Sazili. 2012. Halal authenticity issues in meat and meat products. Meat Sci. 91: 207-214.
Ni’mah, A., Y. Kartikasari, A. D. Pratama, L. R. Kartikasari, B. S. Hertanto, and M. Cahyadi. 2016. Detection of pork contamination in fresh and cooked beef using genetic marker mitochondrial-DNA cytochrome b by duplex-PCR. J. Indonesian Trop. Anim. Agric. 41: 7-12.
Rahmawati, Sismindari, Raharjo, T. J. Sudjadi, and A. Rohman. 2016. Analysis of pork contamination in Abon using mitochondrial D-Loop22 primers using real time polymerase chain reaction method. Int. Food Res. J. 23: 370-374.
Sahilah, A. M., Y. Norhayati, A. S. Norrakiah, A. Aminah, and W. M. Wan Aida. 2011. Halal authentification of raw meats using PCR amplification of mitochondrial DNA. Int. Food Res. J. 18: 1489-1491.
Soares, S., J. S. Amaral, I. Mafra, and M. B. P. P. Oliveira. 2010. Quantitative detection of poultry meat adulteration with pork by a duplex PCR assay. Meat Sci. 85: 531-536.
Sotelo, C. G., P. Calo-Mata, M. J. Chapela, R. I. Pérez-Martín, G. L. Hold, V. J. Russell, S. Pryde, J. Quinteiro, M. Izquierdo, M. Rey-Méndez, C. Rosa, and A. T. Santos. 2001. Identification of Flatfish (Pleuronectiforme) Species Using DNA-Based Techniques. J. Agric. Food Chem. 49: 4562-4569. DOI: 10.1021/jf010452a
Zhang, C. 2013. Semi-nested multiplex PCR enhanced method sensitivity of species detection in further-processed meats. Food Control. 31: 326-330.
DOI: https://doi.org/10.21059/buletinpeternak.v41i2.13376
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