THE USE OF QUANTITATIVE POLYMERASE CHAIN REACTION (qPCR) METHOD FOR PIG DNA DETECTION

  • Bambang Hermawan Institut Pertanian Bogor
  • Riska Dwi Nanda
  • Nadya Nurafifah Andriya
  • Jakaria

Abstract

The use of food ingredients and/or processed food products contaminated with pork has become a growing concern and issue at present. This condition encourages the development of an accurate method for specifically detecting the presence or absence of pork contamination. The aim of this research is to detect pig DNA using the quantitative Polymerase Chain Reaction (qPCR) method. The research was carried out in two stages, namely (1) using fresh meat to detect DNA and cDNA to provide a positive control, and (2) product samples in the form of pork-based preparations (floss, meatballs, corned beef and sausages) were tested using DNA markers to determine robustness of the detection method used. The research stages carried out were DNA extraction and RNA isolation from pork that had been modified and developed. Then proceed to measure the purity and concentration of DNA/RNA using a spectrophotometer. The RNA isolation results were converted into complementary DNA (cDNA) and together with the DNA extraction results were analyzed using qPCR with a specific primer sequence for pig (Sus scrofa) DNA. The results of measuring the extraction concentration showed that the DNA concentration was 71.1296.025 ng/ul with varying purity. DNA qPCR is more time efficient than cDNA qPCR because it does not require the RNA transcription stage. Pork DNA was amplified in all samples compared to the positive control in the Ct range of 23-28 ng/ul, so that pork contamination can be properly detected from the processed food products analyzed

Downloads

Download data is not yet available.

References

Ardilla D, Taufik M, Tarigan DM, Thamrin M, Razali M, Siregar HS. 2018. Analisis lemak babi pada produk pangan olahan menggunakan spektroskopi UV-Vis. Jurnal Teknologi Pangan & Hasil Pertanian. 1(2): 111116.
Cai Y, Li X, Lv R, Yang J, Li J, He Y, Pan L. 2014. Quantitative analysis of pork and chicken products by droplet digital PCR. BioMed Research International. 2014(2014): 810209.
Canggih C, Fikriyah K, Yasin A. 2017. Inklusi pembayaran zakat di Indonesia. Jurnal Ekonomi dan Bisnis Islam. 3(1): 111.
Cawthorn DM, Steinman HA, Witthuhn RC. 2011. Comparative study of different methods for the extraction of DNA from fish. Food Control. 22(2): 231244.
Coyle PV, Al Molawi NH, Kacem MABH, El Kahlout RA, Al Kuwari E, Al Khal A, Gillani I, Jeremijenko A, Saeb H, Al Thani M, Bertollini R, Rahim HFA, Chemaitelly H, Tang P, Latif AN, Al Kaabi S, Al Maslaman MARS, Morris BD, Al-Ansari N, Kaleeckal AH, Raddad LJA. 2022. Reporting of RT- PCR cycle threshold (Ct) values during the first wave of COVID-19 in Qatar improved result interpretation in clinical and public health settings. Journal of Medical Microbiology. 71(5): 16.
Dastgheib S, Irajie C, Assaei R, Koohpeima F, Mokarram P. 2014. Optimization of RNA Extraction from Rat Pancreatic Tissue. Iranian Journal of Medical Science. 39(3): 282288.
Dayanti FG, Djuminar A, Dermawan A, Tanta A. 2019. Perbandingan nilai pengukuran kuantitatif hasil ekstraksi DNA Salmonella typhi menggunakan metode boiling, NaOH, kit komersial. Jurnal Riset Kesehatan. 11(1): 350357.
Indrasti D, Mukhlisin MF, Darmawan N, Yuliana ND. 2022. Profil komponen volatil beberapa jenis satai menggunakan kromatografi gas. Jurnal Ilmu Pertanian Indonesia. 27(2): 199215.
Irwandi, Wardi ES, Dova S. 2020. Deteksi cemaran gen babi pada produk bakso sapi kemasan di kotaPadang menggunakan metode PCR (polymerase chain reaction). Jurnal Akademi Farmasi Prayoga. 5(2): 1021.
Fitrianingsih, Tasse AM, Astuti F, Rahayu N, Sani LOA, Sutopo D, Abad M. 2022. Teknologi pembuatan abon daging sapi untuk meningkatkan pendapatan hasil ternak pasca Covid-19 di desa Kiaea kabupaten Konawe Selatan. Dalam: Prossiding Seminar Nasional Inovasi dan Teknologi Peternakan II: Optimasi Integrated Farming System Berbasis Teknologi Peternakan dalam Menunjang Pemenuhan Protein Hewani di Era New Normal. Kendari, 19 November 2022.
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.
Kuswandi B, Gani AA, Ahmad M. 2017. Immuno strip test for detection of pork adulteration in cooked meatballs. Food Bioscience. 19: 16.
Lam S, Kommadath A, Campos ÓL, Prieto N, Aalhus J, Juárez M, Dugan MER, Vahmani P. 2021. Evaluation of RNA quality and functional transcriptome of beef longissimus thoracis over time post-mortem. PloS ONE. 16(5): 114.
Mariyani, Sismindari, Rumiyati. 2021. Validasi metode real-time polymerase chain reaction untuk deteksi DNA babi (Sus scrofa domestica) dan celeng (Sus barbatus) pada sosis sapi. Jurnal Ilmiah Indonesia. 6(8): 39252940
Mustaqimah DN, Septiani T, Roswien AP. 2020. Deteksi DNA babi pada produk sosis menggunakan real time-polymerase chain reaction (RT-PCR). 3(2): 106111.
Nakyinsige K, Che Man Y, Sazili AQ. 2012. Halal authenticity issues in meat and meat products. Meat Science. 91(3): 207–214.
Nygard AB, Jørgensen CB, Cirera S, Fredholm M. 2007. Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR. BMC Molecular Biology. 8: 67.
Orbayinah S, Hari W, Adam H, Sismindari S, Abdul R. 2019. Application of real-time polymerase chain reaction using species specific primer targeting on mitochondrial cytochrome-b gene for analysis of pork in meatball products. Journal of Advanced Veterinary and Animal Research. 6(2): 260–265.
Pagani S, Maglio M, Sicuro L, Fini M, Giavaresi G, Brogini S. 2023. RNA extraction from cartilage: issues, methods, tips. International Journal of Molecular Sciences. 24: 1–22.
Panglipur PE, Sulandari L. 2014. Pengaruh jumlah salad oil dan CMC (carboxy methyl cellulose) terhadap sifat organoleptik kornet daging sapi. E-Journal Boga. 3(1): 160–165.
Rahmania YL, Widayat, Agustini TW, Suzery M, Albaari AN. 2021. Pengukuran kandungan DNA babi dalam berbagai produk pengan dengan metode real time polymerase chain reaction (RT-PCR). Indonesia Journal of Halal. 3(2): 129–133.
Sign KP, Miaskowski C, Dhruva AA, Flowers E, Kober KM. 2018. Mechanisms and measurement of changes in gene expression. Biological Research for Nursing. 20(4): 369–382.
Stefanova P, Taseva M, Georgieva T, Gotcheva V, Angelov A. 2013. A modified CTAB method for DNA extraction from soybean and meat products. Biotechnology & Biotechnological Equipment. 27(3): 3803–3810.
Waluyo S, Malau J, Raekiansyah M, Yulian E, Hardiman I. 2019. In silico analysis of actin gene as a candidate for DNA non-halal detection base on real-time PCR. Indonesian Journal of Halal Research. 3(2): 70–74.
Widayat, Agustini TW, Suzery M, Al-Baarri AN, Putri SR, Kurdianto. 2019. Real time-polymerase chain reaction (RT-PCR) sebagai alat deteksi DNA babi dalam beberapa produk non-pangan. Indonesian Journal of Halal. 2(1): 26–33.
Yalçınkaya B, Yumbul E, Mozioğlu E, Akgoz M. 2017. Comparison of DNA extraction methods for meat analysis. Food Chemistry. 221: 1253–1257.
Published
2024-07-02
How to Cite
HermawanB., NandaR. D., AndriyaN. N., & Jakaria. (2024). THE USE OF QUANTITATIVE POLYMERASE CHAIN REACTION (qPCR) METHOD FOR PIG DNA DETECTION. Jurnal Ilmu Pertanian Indonesia, (00). Retrieved from https://jurnalpenyuluhan.ipb.ac.id/index.php/JIPI/article/view/50577
Issue
Vol. 0 No. 00 (2024): inpress
Section
Articles
Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

This journal is published under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License. Authors who publish with this journal agree to the following terms: Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. NonCommercial — You may not use the material for commercial purposes.