Detection of species substitution in raw, cooked, and processed meats utilizing multiplex-PCR assay

Muhammad Cahyadi; Nur Aini Dyah Fauzıah; Imam Tubagus Suwarto; Waraporn Boonsupthip

doi:10.22146/ijbiotech.63472

Detection of species substitution in raw, cooked, and processed meats utilizing multiplex-PCR assay

https://doi.org/10.22146/ijbiotech.63472

Muhammad Cahyadi^(1*), Nur Aini Dyah Fauzıah⁽²⁾, Imam Tubagus Suwarto⁽³⁾, Waraporn Boonsupthip⁽⁴⁾

(1) Department of Animal Science, Faculty of Agriculture, Universitas Sebelas Maret, Surakarta 57126, Indonesia; Halal Research Center and Services, Institute of Research and Community Service, Universitas Sebelas Maret, Surakarta 57126, Indonesia
(2) Department of Animal Science, Faculty of Agriculture, Universitas Sebelas Maret, Surakarta 57126, Indonesia
(3) PT. Charoen Pokphand Jaya Farm Mekarsari, Sukabumi 43196, Indonesia
(4) Department of Food Science and Technology, Faculty of Agro‐Industry, Kasetsart University, Bangkok 10900, Thailand
(*) Corresponding Author

Abstract

The rise of beef consumption in Indonesia opens an opportunity for “rogue” suppliers to mix beef with other meat species that are relatively cheaper, such as pork, chicken, etc. The aim of this study was to identify pig and chicken meat in raw, cooked, and processed meat products using multiplex-PCR of mitochondrial DNA Cytochrome b gene, which is maternally inherited and widely used for forensic studies. A total of 90 samples-33 raw meats, 33 cooked meats, and 24 meatballs-were used in this study. Each sample was extracted to obtain the DNA genome and this was then amplified using multiplex-PCR. The PCR products were visualized using 2% agarose gel electrophoresis. The results showed that species contained in raw, cooked, and processed meat samples could be identified as indicated by DNA bands at 398, 274, 227, and 157 bp for pig, cattle, chicken, and goat species respectively. This study concluded that species substitution in raw, cooked, and processed meats could be detected using the Cytochrome b gene as a genetic marker through multiplex-PCR assay.

Keywords

Cytochrome b, meatball, meat substitution, multiplex-PCR

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References

Ali M, Mustafa S, Hashim U, Che Man Y, Foo K. 2012. Nanobioprobe for the determination of pork adulteration in burger formulations. J Nanomater. 2012. doi:10.1155/2012/832387.

Ali ME, Razzak MA, Hamid SBA, Rahman MM, Amin MA, Rashid NRA, Asing. 2015. Multiplex PCR assay for the detection of five meat species forbidden in Islamic foods. Food Chem. 177:214–224. doi:10.1016/j.foodchem.2014.12.098.

Ballin NZ, Vogensen FK, Karlsson AH. 2009. Species determination Can we detect and quantify meat adulteration? Meat Sci. 83(2):165–174. doi:10.1016/j.meatsci.2009.06.003.

Bintoro V. 2008. Teknologi Pengolahan Daging dan Analisis Produk [Bahasa Indonesia]. Semarang: Badan Penerbit Universitas Diponegoro.

Cahyadi M, Wibowo T, Pramono A, Abdurrahman ZH. 2020. A novel multiplexpcr assay to detect three nonhalal meats contained in meatball using mitochondrial 12s rrna gene. Food Sci Anim Resour. 40(4):628–635. doi:10.5851/kosfa.2020.e40.

Castresana J. 2001. Cytochrome b phylogeny and the taxonomy of great apes and mammals. Mol Biol Evol. 18(4):465–471. doi:10.1093/oxfordjournals.molbev.a003825.

Chisholm J, Conyers C, Booth C, Lawley W, Hird H. 2005. The detection of horse and donkey using realtime PCR. Meat Sci. 70(4):727–732. doi:10.1016/j.meatsci.2005.03.009.

Fajardo V, González I, Martín I, Rojas M, Hernández PE, García T, Martín R. 2008. Realtime PCR for quantitative detection of chamois (Rupicapra rupicapra) and pyrenean ibex (Capra pyrenaica) in meat mixtures. J AOAC Int. 91(1):103–111. doi:10.1093/jaoac/91.1.103.

Ghovvati S, Nassiri MR, Mirhoseini SZ, Moussavi AH, Javadmanesh A. 2009. Fraud identification in industrial meat products by multiplex PCR assay. Food Control. 20(8):696–699. doi:10.1016/j.foodcont.2008.09.002.

Haider N, Nabulsi I, AlSafadi B. 2012. Identification of meat species by PCRRFLP of the mitochondrial COI gene. Meat Sci. 90(2):490–493. doi:10.1016/j.meatsci.2011.09.013.

He H, Hong X, Feng Y, Wang Y, Ying J, Liu Q, Qian Y, Zhou X, Wang D. 2015. Application of quadruple multiplex PCR detection for beef, duck, mutton and pork in mixed meat. J Food Nutr. Res. 3(6):392–398. doi:10.12691/jfnr366.

Hossain MA, Ali ME, Hamid SBA, Asing, Mustafa S, Desa MNM, Zaidul ISM. 2017. Targeting double genes in multiplex PCR for discriminating bovine, buffalo and porcine materials in food chain. Food Control. 73:175–184. doi:10.1016/j.foodcont.2016.08.008.

Kesmen Z, Gulluce A, Sahin F, Yetim H. 2009. Identification of meat species by TaqManbased realtime PCR assay. Meat Sci. 82(4):444–449. doi:10.1016/j.meatsci.2009.02.019.

Lockley AK, Bardsley RG. 2000. DNAbased methods for food authentication. Trends Food Sci Technol. 11(2):67–77. doi:10.1016/S09242244(00)000492.

Mane BG, Mendiratta SK, Tiwari AK. 2009. Polymerase chain reaction assay for identification of chicken in meat and meat products. Food Chem. 116(3):806– 810. doi:10.1016/j.foodchem.2009.03.030.

Mane BG, Mendiratta SK, Tiwari AK. 2012. Beef specific polymerase chain reaction assay for authentication of meat and meat products. Food Control. 28(2):246– 249. doi:10.1016/j.foodcont.2012.05.031.

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 Sci. 51(2):143– 148. doi:10.1016/S03091740(98)001120.

Mohsin A, Ramli N, Alkhulayfi BA. 2016. Halal tourism: Emerging opportunities. Tour Manag Perspect. 19:137–143. doi:10.1016/j.tmp.2015.12.010.

Murugaiah C, Noor ZM, Mastakim M, Bilung LM, Selamat J, Radu S. 2009. Meat species identification and Halal authentication analysis using mitochondrial DNA. Meat Sci. 83(1):57–61. doi:10.1016/j.meatsci.2009.03.015.

Nakyinsige K, Man YBC, Sazili AQ. 2012. Halal authenticity issues in meat and meat products. doi:10.1016/j.meatsci.2012.02.015.

Novianty E, Kartikasari LR, Lee JH, Cahyadi M. 2017. Identification of pork contamination in meatball using genetic marker mitochondrial DNA cytochrome b gene by duplexPCR. IOP Conf Ser: Mater Sci Eng. 193(1). doi:10.1088/1757899X/193/1/012002.

Rohman A, Sismindari, Erwanto Y, Che Man YB. 2011. Analysis of pork adulteration in beef meatball using Fourier transform infrared (FTIR) spectroscopy. Meat Sci. 88(1):91–95. doi:10.1016/j.meatsci.2010.12.007.

van der Spiegel M, van der FelsKlerx HJ, Sterrenburg P, van Ruth SM, ScholtensToma IM, Kok EJ. 2012. Halal assurance in food supply chains: Verification of halal certificates using audits and laboratory analysis. Trends Food Sci Technol. 27(2):109–119. doi:10.1016/j.tifs.2012.04.005.

DOI: https://doi.org/10.22146/ijbiotech.63472