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Submitted
Mar 3, 2019
Accepted
Apr 2, 2019
Published
Mar 31, 2019
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Abstract

The frequency of non-halal ingredient mixing, such as porkin on the food processed products of meatballs, has become an issue to the public, especially for moslems. Therefore, a reliable and valid method with high sensitivity is needed to specifically detect the pig contamination. This research aims to obtain a valid and reliable method by proposing polymerase chain reaction (PCR)  using mitochondrial D-Loop22 primer as a method in handing halal food authentication. The sample consisted of beef as a negative control, pork and pork meatballs as a positivecontrol, and five samples of meat balls found in Makassar for halal inspection.The method validation assay was conducted by testing the primary specificity on the fresh tissue (beef and pork) and testing the sensitivity by making a series of pig DNA dilution (1:10;  1:102;   1:103;   1:104)   and the variations of contaminated pig:cow (%b/b) : 0.05% , 0.1%, 1%, and 5%. The result of PCR amplification on agarose gel electrophoresis of 0.8% showed that method was able to detect the pig DNA contamination specifically in pigs and not amplify other DNA, and could still be detected up to pig contamination specifically in pigs and not amplify the other DNAs and could still be detected up to pig contamination of 0.05% and on DNA dilution of 1:103. Meanwhile, on the five samples analyzed, there were  not found pig DNA contamination characterized by no formed amplification bands.

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How to Cite
Wahyuni, S., Maryam, S., & Aminah, A. (2019). Validasi Metode Analisis Cemaran DNA Babi pada Bakso Sapi Menggunakan Primer Mitokondria D-Loop22 dengan Metode Polymerase Chain Reaction (PCR): Analysis Method Validation of Pig DNA Contamination in Cow Meatballs Using Mitochondrial Primer D-Loop22 by Polymerase Chain Reaction (PCR) Method. Jurnal Farmasi Galenika (Galenika Journal of Pharmacy) (e-Journal), 5(1), 65 - 72. https://doi.org/10.22487/j24428744.2019.v5.i1.12035

References

  1. Ardi, A. (2012). Validasi metode ekstraksi DNA pada analisis DNA babi dalam produk bakso, [Skripsi]. Fakultas MIPA. Institut Pertanian Bogor. Bogor.

    Farmawati, D.A., Wirajana, N., & Yowani, S.C. (2015). Perbandingan Kualitas DNA dengan menggunakan Metode Boom Original dan Boom Modifikasi pada Isolat Mycobacterium tuberculosis 151. KIMIA, vol. q, pp. 41-45.

    Fatchiyah,. Aruminingtyas, E.L., Widyarti, S., & Rahayu, S. (2011). Biologi Molekuler-Prinsip Dasar Analisis. Erlangga, Malang.

    Fatimah, S. (2013). Deteksi Cemaran Daging Babi dalam Campuran Bakso Ayam dengan Real-Time PCR dan Spektrofotometri FTIR, [Tesis], Fakultas Farmasi, Universitas Gadjah Mada, Yogyakarta.

    Fauziah, A.R. (2016). Identifikasi Cemaran Kulit Babi pada Kerupuk Rambak dengan Metode Real-Time Polymerase Chain Reaction menggunakan Primer Mitokondria D-Loop 22', [Skripsi], Fakultas Farmasi, Universitas Gadjah Mada, Yogyakarta.

    Hertanto, B.S., Fitra, R.A., Kartikasari, L.R., & Cahyadi, M. (2017). Authentication of Raw Chicken Mear from Pork Contamination using Gene CYT-B with Duplex-Polymerase Chain Reaction Analysis, Buletin Peternakan, vol. 41, pp. 113–118

    Marlina, Mutalib, S.A., Islami, Sari, H.K., & Fitria, A. (2013). Pengembangan Metode PCR dan Southern Hybridization untuk Deteksi Gen Babi pada Cangkang Kapsul. Pros. Semin. Nas. Perkemb. Terkini Sains Farm. Dan Klin. III, pp. 116–121

    Maryam, S. (2014). Analisis Cemaran DNA Babi Pada Dendeng menggunakan primer mitokondria D-Loop686 dan gen cyt b dengan menggunakan Real time polymerase chain reaction (real time PCR), [Tesis], Fakultas Farmasi. Universitas Gadjah Mada, Yogyakarta.

    Maryam, S., Sismindari, Raharjo, T.J., Sudjadi, Rohman, A. (2015). Determination of Porcine Contamination in Laboratory Prepared dendeng Using Mitochondrial D-Loop686 and cyt b Gene Primers by Real Time Polymerase Chain Reaction, International Journal of Food Properties, vol.19, no. 11, pp. 187–195. 2017,https://doi.org/10.1080/10942912.2015.1020434

    Nugroho, K., Terryana, R.T., dan Lestari, P. (2017). Metode Ekstraksi DNA Cabai (Capsicum annuum L.) menggunakan Modifikasi Buffer CTAB (Cethyl Trimethyl Ammonium Bromide) tanpa Nitrogen Cair, Scripta Biologi, vol. 4, no. 2, pp. 91-94

    Raharjo, T.J., Alfiraza, E.N., Enjelina, E., & Pranowo, D. (2017). Validation of a Non-Specific Dye Real-Time PCR Assay for Porcine Adulteration in Meatball Using ND5 Primer,  Indones. J. Chem, vol. 17, no.2, pp. 167–174.

    Rahmawati, Sisimindari, Raharjo, T.J., Sudjadi, & Rohman, A. (2016). Analysis of Pork Contamination in Abon using Mitochondrial D-Loop 22 primer Using Real-Time Polymerase Chain Reaction Method, International Food Research Journal, vol. 23, pp. 370-374.

    Rohman, A., Himawati, A., Triyana, K., Sismindari, & Fatimah, S. (2017. 'Identification of Pork in Beef Meatballs using Fourier Transform Infrared Spectrophotometry and Real-time Polymerase Chain Reaction, International Journal of Food Properties, vol. 20, pp. 654-661.

    Sambrook, J., Fritsch, E.F., & Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, pp. 626.

    Sugihartini, N., Fudholi, A., Pramono, S., & Sismindari, S. (2014). Validasi Metode Analisa Penetapan Kadar Epigalokatekin Galat dengan Kromatografi Cair Kinerja Tinggi, Pharmaciana, vol. 4, no. 1, pp. 39–44.

    Tenriulo, A., Suryati, E., Parenrengi, A., & Rosmiati. (2001). Ekstraksi DNA Rumput Laut Rappaphycus alvarezil dengan metode Fenol Kloroform’, Marina Chimica Acta, vol. 2, no. 2, pp. 6-10

    Zulfahmi. (2015). Deteksi Kontaminan Babi Pada Produk Makanan Menggunakan Teknologi DNA Molekuler, Kutubkhanah: J. Penelit. Sos. Keagamaan, vol. 18, no. 1, pp.