Detection of Porcine Contamination in Beef Meatballs from Gedangan District Using PCR–RFLP
DOI:
https://doi.org/10.15575/biodjati.v10i2.49813Keywords:
BsaJI, detection food halal, meatball, PCR-RFLPAbstract
Meatball is one of the popular meat dishes in Indonesia, especially in Sidoarjo. The sale of meatballs in Sidoarjo is carried out by Micro, Small, and Medium Enterprises (MSMEs), with 330 of the total 11,114 micro-traders selling meatballs. However, there is a risk of pork being adulterated into meatballs. This is a serious problem because Indonesia, especially Sidoarjo, is a Muslim-majority city with 95.5% of the total population of 2 million. Therefore, the purpose of this article is to conduct field checks to determine whether or not pork contamination is present in meatballs in Sidoarjo. Sampling locations were determined using a stratified random sampling method, which found that eight of the 15 villages, including Karangbong, Tebel, Sruni, Keboananom, Keboansikep, Gedangan, Ketajen, and Sawotratap Villages, have a significance value greater than the alpha value (0.05). Therefore, eight of them were used as sampling locations. One sample was purchased from each of the eight chosen villages, named S1, S2, S3, S4, S5, S6, S7, and S8. Meanwhile, the possibility of porcine contamination in beef meatball samples was analyzed using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) method with the restriction enzyme BsaJI. This study found that all eight samples and the negative control DNA amplicons were not cleaved by the BsaJI enzyme. Meanwhile, the positive control DNA amplicon was cleaved into 3 fragments. It was indicated that all samples and the negative control DNA amplicons were free of pork DNA contamination. Despite the anomaly, it can be concluded that all meatball samples bought from Gedangan District are not contaminated with pork. By demonstrating that meatballs from Sidoarjo are free from pork DNA, this study serves as an additional halal food monitoring in Sidoarjo. It reinforces public trust in local MSME food products.
References
Abidin, Z. (2016). Tiga Penjual Daging Nekat Ngecer Daging Celeng dengan Label Daging Sapi. Suara Surabaya.
Abraham, O. S. J., Miguel, T. S., Inocencio, H. C., & Blondy, C. C. (2017). A quick and effective in-house method of DNA purification from agarose gel, suitable for sequencing. 3 Biotech, 7, 180. DOI 10.1007/s13205-017-0851-1.
Adiningsih, M. W., Soejoedono, R. D., Purnawarman, T., Latif, H., Adji, R. S., Poetri, O. N., & Putri, D. D. (2018). Identification of sumateran wild boar meat (Sus scrofa vittatus) by Restriction fragment length polymorphism (RFLP) analysis of cytochrome b gene. In Proc. of the 20th FAVA CONGRESS & The 15th KIVNAS PDHI, Bali. Retrieved from: https://www.academia.edu/download/114521548/15552.pdf
Azad, M. A. K., Dey, M., Khanam, F., Biswas, B., & Akhter, S. (2023). Authentication of meat and meat products using molecular assays: A review. Journal of Agriculture and Food Research, 12, 100586. DOI: 10.1016/j.jafr.2023.100586.
BPOM. (2018). Peraturan Badan Pengawas Obat dan Makanan Nomor 31 Tahun 2018 tentang Label Pangan Olahan. Jakarta: Badan Pengawas Obat dan Makanan.
BPS. (2024). Jumlah penduduk menurut kecamatan dan agama yang dianut (jiwa), 2023. Badan Pusat Statistik Kabupaten Sidoarjo.
BPS Statistics Sidoarjo Regency. (2024). Gedangan District in Figures 2024. Sidoarjo: BPS Statistics Sidoarjo Regency.
Bryan, T. M., & Baumann, P. (2011). G-quadruplexes: from guanine gels to chemotherapeutics. Molecular biotechnology, 49, pp. 198-208. DOI: 10.1007/s12033-011-9395-5
Cozzaglio, M., Bimbo, L., Armas, P., & Lago, S. (2022). G-quadruplex formation within the promoter of TEAD4: Biophysical and cellular evidence. Frontiers in Chemistry, 10, 1008075. DOI: 10.3389/fchem.2022.1008075
Das, P. J., Kumar, S., Choudhury, M., Banik, S., Pegu, S. R., Kumar, S., Deb, R., & Gupta, V. K. (2024). Characterization of the complete mitochondrial genome and identification of signature sequence of Indian wild pig. Gene, 897, 148070. DOI: 10.1016/j.gene.2023.148070
Díaz, T., Ortega-Pinazo, J., Martínez, B., Jiménez, A., Gómez-Zumaquero, J. M., Caracuel, Z., Hortas, M. L., & Ferro, P. (2023). Measurement of yield and quality of DNA in human buffy coat is extraction method dependent. Preparative Biochemistry & Biotechnology, 53(6), pp. 591-598. DOI: 10.1080/10826068.2022.2119574
Ditakopum. (2025). Data umum usaha mikro Kabupaten Sidoarjo. Dinas Koperasi dan Usaha Mikro Kabupaten Sidoarjo. Retrieved from: https://ditakopum.sidoarjokab.go.id/public/usaha-mikro (Accessed: 05 July 2025).
Elyasigorji, Z., Izadpanah, M., Hadi, F., & Zare, M. (2023). Mitochondrial genes as strong molecular markers for species identification. The Nucleus, 66(1), pp. 81-93. DOI: 10.1007/s13237-022-00393-4
Erwanto, Y., Abidin, M. Z., Muslim, E. Y. P., Sugiyono, & Rohman, A. (2014). Identification of pork contamination in meatballs of Indonesia local market using polymerase chain reaction restriction fragment length polymorphism (PCR RFLP) analysis. Asian Australasian Journal of Animal Sciences, 27(10), 1487–1492. DOI: 10.5713/ajas.2014.14014
Erwanto, Y., Sugiyono, & Rohman, A. (2013). Identification of pork contamination in meatballs of indonesia local market using polymerase chain reaction restriction fragment length polymorphism (PCR RFLP) Analysis. agriTECH, 33(2), 123–129. DOI: 10.22146/agritech.9579
Fadhlurrahman, Wardani, A. K., & Widyastuti, E. (2015). Deteksi gelatin babi pada soft candy menggunakan metode PCR RFLP sebagai salah satu pembuktian kehalalan pangan. Jurnal Teknologi Pertanian, 16(2). https://jtp.ub.ac.id/index.php/jtp/article/view/490/0
Farag, W. M. H., Aljuaydi, S. H., Galal, M. K., Kassem, G. M. A., & Gouda, E. M. (2022). Efficient Mitochondrial Genes in the Characterization of Meat Species Applying PCR RFLP Technique. Advances in Animal and Veterinary Sciences, 10(1): 8-13. DOI: 10.17582/journal.aavs/2022/10.1.8.13
Fatima, S., Javed, M., Zahra, H., Hussain, I., & Mian, A. A. (2024). A Rapid, cost effective method for high yield dna purification from PCR products and agarose gels. bioRxiv, 2024-11. DOI: 10.1101/2024.11.13.623516
Finnis, J., Murphy, C., Davidson, G., Alexander, K., Lewis, J., Boyce, M., Kennedy, F., Casey, D., Clayson, N., Fraser, I., Hargreaves, C., Stevenson, N., Doole, S., Rogers, C. (2023). Enzyme activity, DNA degradation and drying times of semen, saliva and vaginal material. Science & Justice, 63(6): pp. 663 – 670. DOI: 10.1016/j.scijus.2023.09.001
Fitriani, L. N. (2023). Deteksi DNA babi (Sus scrofa) pada produk kornet dengan metode polymerase chain reaction fragment length polymorphism (PCR-RFLP), Skripsi, Sarjana Ilmu Pangan dan Bioteknologi, Fakultas Teknologi Pangan, Universitas Brawijaya.
Government of Sidoarjo Regency. (2025). Circular letter of the regent of sidoarjo on halal certification to support halal tourism destinations in Sidoarjo [Circular Letter]. Sidoarjo Regency Communication and Information Office.
Griffiths, K. R., McLaughlin, J. L. H., Hall, F., Partis, L., Hansen, S. C., Tulloch, R., & Burke, D. G. (2023). Development of Seven New dPCR Animal Species Assays and a Reference Material to Support Quantitative Ratio Measurements of Food and Feed Products. Foods, 12(20), 3839. DOI: 10.3390/foods12203839
Grün, J. T., & Schwalbe, H. (2021). Folding dynamics of polymorphic G-quadruplex structures. Biopolymers, 113(1), e23477. DOI: 10.1002/bip.23477
Han, X., Xie, X., Zhao, R., Li, R., Ma, P., Li, H., Chen, F., Zhao, Y., & Tang, Z. (2024). Calculating the similarity between prescriptions to find their new indications based on graph neural network. Chinese Medicine. DOI: 10.1186/s13020-024-00994-y
Hardianto, A., Kusumaningtyas, P., Prabowo, S., & Hartati, Y. W. (2023). DNA based detection of pork content in food. Heliyon, 9(3). DOI: 10.1016/j.heliyon.2023.e14418
Hays, A., Islam, R., Matys, K., & Williams, D. (2022). Best Practices in qPCR and dPCR Validation in Regulated Bioanalytical Laboratories. The AAPS Journal, 24: 36. DOI: 10.1208/s12248-022-00686-1
Henderson, E., Hardin, C. C., Walk, S. K., Tinoco Jr, I., & Blackburn, E. H. (1987). Telomeric DNA oligonucleotides form novel intramolecular structures containing guanine-guanine base pairs. Cell, 51, pp. 899–908. DOI: 10.1016/0092-8674(87)90577-0
Irwandi, I., Wardi, E. S., & Dova, S. (2020). Deteksi cemaran gen babi pada produk bakso sapi kemasan di Kota Padang menggunakan metode PCR (polymerase chain reaction). Jurnal Akademi Farmasi Prayoga, 5(2), pp. 10-22. DOI: 10.56350/jafp.v5i2.63
Jung, S.-J., Lee, J. H., Lim, J.-Y., & Yang, Y.-Y. (2024). Mitochondrial DNA Copy Number Is Associated with the Severity of Irritable Bowel Syndrome. Medicina, 60(10), 1605. DOI: 10.3390/medicina60101605
Kementerian Agama Republik Indonesia. (2024). Jumlah Penduduk Menurut Agama. Jakarta. Retrieved from: https://satudata.kemenag.go.id/dataset/detail/jumlah-penduduk-menurut-agama (Accessed: 29 May 2024).
Khaira, A., Achyar, A., Atifah, Y., Putri, D. H., & Violita. (2023). Primer Design and Optimization of Annealing Temperature for Analysis of Glutathione Reductase Gene Expression in Rice (Oryza sativa L.). 3BIO: Journal of Biological Science, Technology and Management, 5(1): 142-148. DOI: 10.5614/3bio.2025.7.2
Khamim, M. T. A., & Qurrohman, M. T. (2024). Comparison of DNA isolation of Candida albicans with filter-based kit and cooling methods. International Journal of Health Science and Technology, 5(2), 142-148. DOI: 10.31101/ijhst.v5i2.3273
Khan, A., Carter, R., Mpamhanga, C. D., Masiga, D., Channumsin, M., Ciosi, M., Manangwa, O., Mramba, F., Ijaz, U. Z., Auty, H., Mable, B. K. (2025). Swatting Flies: Biting Insects as Non-Invasive Samplers for Mammalian Population Genomics. Molecular Ecology, e17661. DOI: 10.1111/mec.17661
Khehra, N., Padda, I. S., Zubair, M., (2025). Polymerase Chain Reaction (PCR). StatPearls Publishing LLC.
Kim, S. H., Lee, S. Y., Kim, U., & Oh, S. W. (2023). Diverse methods of reducing and confirming false-positive results of loop-mediated isothermal amplification assays: A review. Analytica Chimica Acta, 1280, 341693. DOI: 10.1016/j.aca.2023.341693
Latham, S., Hughes, E., Budgen, B., Morley, A. (2023). Inhibition of the PCR by genomic DNA. PLoS One, 18(4):e0284538. DOI: 10.1371/journal.pone.0284538
LPPOM MUI. (2024). Wajib Halal, Ini 3 Langkah LPPOM MUI Bantu Pedagang Kaki Lima. Lembaga Pengkajian Pangan, Obat obatan, dan Kosmetika Majelis Ulama Indonesia, Bogor.
Luo, Y., Zivkovic, M. L., Wang, J., Rynes, J., Foldynová Trantírkova, S., Trantirek, L., Verga, D., & Mergny, J.L. (2024). A sodium/potassium switch for G4-prone G/C-rich sequences. Nucleic Acids Research, 52(1), 448–461. DOI: 10.1093/nar/gkad1073
Lv, L., Wu, S., Zhang, Y., & Chen, C. (2022). Characterization of G-quadruplexes in the Enterovirus A71 genome. Microbiology Spectrum, 10(4), e00460-22. DOI: 10.1128/spectrum.00460-22
Masnaini, M., Achyar, A., Chatri, M., Putri, D. H., & Ahda Y. (2023). Primer design and optimization of PCR methods for detecting mixed rat meat in food samples. In Proceedings of the 3rd International Conference on Biology, Science and Education (IcoBioSE 2021) (Vol. 32, pp. 282). Springer Nature. https://books.google.co.id/books?hl=en&lr=&id=4d2_EAAAQBAJ&oi=fnd&pg=PA282&dq=Masnaini,+M.,+Achyar,+A.,+Chatri,+M.,+Putri,+D.+H.,+%26+Ahda,+Y.+(2023).+Primer+Design+and+Optimization+of+PCR+Methods+for+Detecting+Mixed+Rat+Meat+in+Food+Samples.+In+Proceedings+of+the+3rd+International+Conference+on+Biology,+Science+and+Education+(IcoBioSE+2021)+(Vol.+32,+pp.+282).+Springer+Nature.+&ots=WugkkqRkQo&sig=462fxnGxCoLa0quSx7_uNlJnEWU&redir_esc=y#v=onepage&q&f=false
Muflihah, Hardianto, A., Kusumaningtyas, P., Prabowo, S., Hartati, Y. W. (2023). DNA based detection of pork content in food. Heliyon, e14418. DOI: 10.1016/j.heliyon.2023.e14418
Muhammed, M. A., Bindu, B. S. C., Jini, R., Prashanth, K. H., & Bhaskar, N. (2015). Evaluation of different DNA extraction methods for the detection of adulteration in raw and processed meat through polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Journal of Food Science and Technology, 52, pp. 514-520. DOI: 10.1007/s13197-013-1024-9
Murphy, Z., Shields, D. A., Evrony, G. D. (2024). Serial Enrichment of Heteroduplex DNA using a MutS magnetic bead system. Biotechnology Journal, 18(1):e2200323. DOI: 10.1002/biot.202200323
Park, D., Kim, Y., Lee, J., & Cho, S. (2024). G.quadruplexes as structural elements influencing replication origin selection. Nature Communications, 15, 6419. DOI: 10.1038/s41467-024-51797-6
Ramlan, R. R., Harnelly, E., Fitri, L. (2024). DNA Extraction and PCR Optimization of Coffea arabica L. and Coffea canephora Pierre ex A. Froehner. Jurnal Penelitian Pendidikan IPA, 10. DOI: 10.29303/jppipa.v10iSpecialIssue.7881
Rani, W. M., Puspitam, R. D., Sefina, N., Sa’adah, N., & Achyar, A. (2024). Analisis variasi genetik gen L1 HPV-52 menggunakan rflp secara in silico. Indonesian Journal of Pharmaceutical Education, 4(1). DOI: 10.37311/ijpe.v4i1.24496
Republic of Indonesia. (2021). Government regulation of the Republic of Indonesia number 39 of 2021 on implementation in the field of halal product assurance (State gazette of the Republic of Indonesia year 2021 no. 49; Supplement to the state gazette no. 6651). Jakarta: State Secretariat. Retrieved July 7, 2025, from https://peraturan.bpk.go.id/Details/161927/pp-no-39-tahun-2021
Rohman, M. S. (2025). Mengejutkan, 88 Persen Daging di Pasar Sidoarjo Ternyata Gelonggongan. Radar Sidoarjo. Retrieved October 22, 2025, from https://radarsidoarjo.jawapos.com/kota-delta/855605734/mengejutkan-88-persen-daging-di-pasar-sidoarjo-ternyata-gelonggongan?.
Rosalinda, E., Sari, A. P. Z. N. L., Saraswati, Y. V., Istarisa, A. F., Sasongko, H., Alfiyanto, A. R., & Maharani, D. (2024). Single nucleotide polymorphism and restriction enzyme analysis of chicken’s prolactin gene based on genbank sequences. In IOP Conference Series: Earth and Environmental Science (Vol. 1360, No. 1, p. 012024). IOP Publishing. DOI: 10.1088/1755-1315/1360/1/012024
Sen, D., & Gilbert, W. (1988). Formation of parallel four stranded complexes by guanine-rich motifs in DNA and its implications for meiosis. Nature, 334, 364–366. DOI: 10.1038/334364a0
Singh, Y., Anand, K. J., Singh, M., Ambawat, S., & Tiwari, G. (2023). Recent developments in PCR technology. Advance in Biological Sciences and Biotechnology, page 77-95, pp. 79. DOI: 10.22271/int.book.316
SISKAPERBAPO. (2024). Sistem Informasi Ketersediaan dan Perkembangan Harga Bahan Pokok di Jawa Timur: Harga Rata Rata Provinsi Jawa Timur di Tingkat Konsumen Tanggal 2024-12-16. Available at: https://siskaperbapo.jatimprov.go.id/harga/tabel (Accessed: 16 December 2024).
Siswara, H. N., Erwanto, Y., & Suryanto, E. (2022). Study of meat species adulteration in Indonesian commercial beef meatballs related to halal law implementation. Frontiers in Sustainable Food Systems, 6, 882031. DOI: 10.3389/fsufs.2022.882031
Sumargo, B. (2020). Teknik sampling. Jakarta: UNJ Press. https://books.google.com/books?hl=en&lr=&id=FuUKEAAAQBAJ&oi=fnd&pg=PA1&dq=Sumargo,+B.+2020.+Teknik+sampling.+Jakarta:+Unj+press.+&ots=nzYe4M-r8L&sig=c4JfQTlV1EIurad80EyDDpqavc0
Sun, H. (2025). Optimization and practical evaluation of multiplex PCR technology for toxin detection in food science. Theoretical and Natural Science, 94, 63–66. DOI: 10.54254/2753-8818/2025.21679
Sundquist, W. I., & Klug, A. (1989). Telomeric DNA dimerizes by formation of guanine tetrads between hairpin loops. Nature, 342, pp. 825–829. DOI: 10.1038/342825a0
Syafrida, S. (2016). Sertifikat halal pada produk makanan dan minumam memberi perlindungan dan kepastian hukum hak-hak konsumen muslim. ADIL: Jurnal Hukum, 7(2): 159-174. DOI: 10.33476/ajl.v7i2.353
Versmessen, N., Simaey, L. V., Negash, A. A., Vandekerckhove, M., Hulpiau, P., Vaneechoutte, M., Cools, P. (2024). Comparison of DeNovix, NanoDrop and Qubit for DNA quantification and impurity detection of bacterial DNA extracts. PLOS One, 19(6): e0305650. DOI: 10.1371/journal.pone.0305650
Wang, K., Jiang, Y., Guo, Y., Geng, M., & Wu, W. (2024). An optimized thermal feedback methodology for accurate temperature control and high amplification efficiency during fluorescent qPCR. Bioengineering, 9(6), 237. DOI: 10.3390/bioengineering9060237
Warmt, C., Yaslanmaz, C., & Henkel, J. (2022). Investigation and validation of labelling loop mediated isothermal amplification (LAMP) products with different nucleotide modifications for various downstream analysis. Scientific Reports, 12, 7137. DOI: 10.1038/s41598-022-11320-7
Yunita, R., Rosadi, F. N., Jamsari, N., & Azizah, A. (2023). Optimization of annealing temperature for amplification of the exon one region of the HPPD gene in HA1 accession sunflowers. IOP Conference Series: Earth and Environmental Science, 1160(1), 012002. DOI: 10.1088/1755-1315/1160/1/012002
Zhang, Y., Li, Y., Lan, W., Hu, X., Yn, S., Liu, Z., Xue, H., Cao, C. (2024). Stable G quadruplex formed in promoter region of oncogene RET in physiological solution conditions. Cell Reports Physical Science, 5(10), 101858. DOI: 10.1016/j.xcrp.2024.101858
Zhou, Y., Bo, F., Tian, T., Wu, B., Zhu, B. (2023). Excessive addition split peak formed by the non-templated nucleotide addition property of Taq DNA polymerase after PCR amplification. Frontier in Bioengineering and Biotechnology, 11. DOI: 10.3389/fbioe.2023.1180542
Zrimec, J., Rijavec, T., Zrimec, T., & Lapanje, A. (2013). Band smearing of PCR amplified bacterial 16S rRNA genes: dependence on initial PCR target diversity. Journal of microbiological methods, 95(2), pp. 186-194. DOI: 10.1016/j.mimet.2013.08.002
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