Specific Species DNA Amplification as a Halal Authentication Method in Sausage Based on Genetic Markers

Nurul Purnomo, Musdalifa Mansur, Angga Nugraha, Muh. Ihsan A. Dagong, Asmuddin Natsir

Abstract


ABSTRACT. Sausage is a processed meat food that is susceptible to counterfeiting by mixing non-halal meat, so a valid, fast and cheap halal authentication method is needed for sausages. This study aims to determine the specific species genes of Cytochrome-b in target DNA amplification as a halal authentication method in sausages. In this study, the samples used were beef sausage, chicken sausage, and pork sausage. The stages of this research consisted of DNA purification, measuring DNA quality, amplifying object DNA using Cytochrome-b, electrophoresis, and visualization of the gel documentation. The DNA purification results from beef sausage, chicken sausage, and pork sausage respectively obtained concentrations of 3 (ng/µl), 2.6 (ng/µl), and 2.8 (ng/µl), and purity of 1.15, 0.76, and 0.88. Meanwhile, DNA amplification produced fragments with lengths for beef sausage, chicken sausage, and pork sausage, namely the Cytochrome-b gene 274 bp, 227 bp, and 398 bp. Based on this study, it's concluded that the Cytochrome-b genes could amplify target DNA from beef sausage, chicken sausage, and pork sausage so that they could be used as gene markers for authenticating halal sausages. 

 

(Amplifikasi DNA Spesifik Spesies sebagai metode autentikasi halal pada sosis berdasarkan penanda genetic) 

ABSTRAK. Sosis merupakan makanan olahan daging yang rentan terhadap pemalsuan dengan mencampurkan daging non halal, sehingga diperlukan metode autentikasi halal yang valid, cepat dan murah untuk sosis. Penelitian ini bertujuan untuk mengetahui gen spesies spesifik Cytochrome-b pada amplifikasi DNA target sebagai metode autentikasi halal pada sosis. Dalam penelitian ini, sampel yang digunakan adalah sosis sapi, sosis ayam, dan sosis babi. Tahapan penelitian ini terdiri dari pemurnian DNA menggunakan Quick-DNA™ Plus Kits, Zymo Research, pengukuran kualitas DNA, amplifikasi pada DNA target menggunakan gen Cytochrome-b, elektroforesis, dan visualisasi pada gel dokumentasi. Hasil pemurnian DNA sosis sapi, sosis ayam, dan sosis babi masing-masing diperoleh konsentrasi 3 (ng/µl), 2,6 (ng/µl), dan 2,8 (ng/µl), dan kemurnian 1,15, 0,76, dan 0,88. Sedangkan amplifikasi DNA menghasilkan fragmen dengan panjang untuk sosis sapi, sosis ayam, dan sosis babi yaitu gen Cytochrome-b 274 bp, 227 bp, dan 398 bp. Berdasarkan penelitian ini, disimpulkan bahwa gen Cytochrome-b dapat mengamplifikasi DNA target dari sosis sapi, sosis ayam, dan sosis babi sehingga dapat digunakan sebagai penanda gen untuk autentikasi sosis halal.


Keywords


gen penanda; halal food; halal sausage; makanan halal; marker gene; olahan daging; PCR; processed meat; sosis halal

Full Text:

PDF

References


Akbar, A., Shakeel, M., Al-amad, S., Akbar, A., Ali, A. K., Rahmeh, R., Alotaibi, M., Al-muqatea, S., Areeba, S., Arif, A., Khan, I. A., Ahmed, S., Hussain, A., & Musharraf, S. G. (2021). A simple and sensitive NGS-based method for pork detection in complex food samples. Arabian Journal of Chemistry, 14(5), 103124. https://doi.org/10.1016/j.arabjc.2021.103124

Arini, R. L., Ramadhani, D., Pebriyanti, N., & Rohman, A. (2018). The use of species-specific primer targeting on D-loop mitochondrial for identification of wild boar meat in meatball formulation. Journal of Advanced Veterinary and Animal Research, 7710(September), 361–368.

Balia, R. L., Suryaningsih, L., & Putranto, W. S. (2014). Pengujian Pemalsuan Bakso dengan Daging Babi Melalui Pendekatan Ensimatis Dan Molekuler Pada UKM di Kawasan Pendidikan Jatinangor Kabupaten Sumedang. Jurnal Aplikasi Ipteks Untuk Masyarakat, 3(2), 70–72.

Barakat, H., El-garhy, H. A. S., & Moustafa, M. M. A. (2014). Detection of pork adulteration in processed meat by species-specific PCR- QIAxcel procedure based on D-loop and cytb genes Detection of pork adulteration in processed meat by species-specific PCR-QIAxcel procedure based on D-loop and cytb genes. Applied Microbiology and Biotechnology, 98, 9805–9816. https://doi.org/10.1007/s00253-014-6084-x

Biase, F. H., Franco, M. M., Goulart, L. R., & Antunes, R. C. (2002). Protocol for extraction of genomic DNA from swine solid tissues. Genetics and Molecular Biology, 25(3), 313–315. https://doi.org/10.1590/S1415-47572002000300011

BPS. (2021). Berita resmi statistik : Hasil Sensus Penduduk 2020. In Bps.Go.Id (Issue 27). https://papua.bps.go.id/pressrelease/2018/05/07/336/indeks-pembangunan-manusia-provinsi-papua-tahun-2017.html

BSN. (2015). Standar Nasional Indonesia Sosis daging. Badan Standar Nasional.

Cahyaningsari, D., Latif, H., & Sudarnika, E. (2019). Identifikasi Penambahan Daging Babi pada Pangan Berbahan Dasar Daging Sapi Menggunakan ELISA dan qPCR. Acta VETERINARIA Indonesiana, 7(2), 17–25. https://doi.org/10.29244/avi.7.2.17-25

Fitrilia, B., Deswinar, D., & Raihana, N. (2017). Deteksi kandungan gen sitokrom B (Cyt b) babi pada bakso yang beredar di Kota Jambi menggunakan teknik Polymerase Chain Reaction. Scientia Jurnal, 6(01), 21–25.

Giasuddin, A. S. M. (1995). Polymerase Chain Reaction Technique: Fundamental Aspects and Applications in Clinical Diagnostics. Journal of Islamic Academy of Sciences, 8(1), 29–32.

Hebert, P. D. N., Cywinska, A., Ball, S. L., & Jeremy, R. (2003). Biological identification through DNA barcodes Biological identifications through DNA barcodes. The Royal Society, 270(January), 313–321. https://doi.org/10.1098/rspb.2002.2218

Hermanto, S., & Meutia, C. D. K. (2009). Perbedaan Profil Protein Produk Olahan (Sosis) Daging Babi dan Sapi Hasil Analisa SDS-PAGE. Jurnal Kimia VALENSI, 1(4), 181–186. https://doi.org/10.15408/jkv.v1i4.247

Hidayat, A. (2020, August). Pasar makanan olahan beku punya prospek positif selama pandemi corona. Kontan.Co.Id, online. https://industri.kontan.co.id/news/pasar-makanan-olahan-beku-punya-prospek-positif-selama-pandemi-corona

Hossain, M. A. M., Muhammad, S., Uddin, K., Sultana, S., Wahab, Y. A., Sagadevan, S., Johan, M. R., Wahab, Y. A., Sagadevan, S., Johan, M. R., & Ali, E. (2020). Authentication of Halal and Kosher meat and meat products : Analytical approaches , current progresses and future prospects. Critical Reviews in Food Science and Nutrition, 0(0), 1–26. https://doi.org/10.1080/10408398.2020.1814691

Hutami, R., Bisyri, H., Nuraini, H., & Ranasasmita, R. (2018). Ekstraksi DNA dari Daging Segar untuk Analisis dengan Metode Loop-Mediated Isothermal Amplification (LAMP) DNA Extraction from Raw Meat for Analysis with the Loop-Mediated Isothermal Amplification (LAMP) Method. Jurnal Agroindustri Halal ISSN 2442-3548, 4(2), 209–216.

Hutami, R., Suprayatmi, M., Idzni, N., Ranasasmita, R., & Nuraini, H. (2018). Study of the Modification of Loop-mediated Isothermal Amplification ( LAMP ) using Taq Polymerase for Halal Testing Study of the Modification of Loop-mediated Isothermal Amplification ( LAMP ) using Taq Polymerase for Halal Testing. January. https://doi.org/10.5220/0009940721912198

Joshi, M., & Deshpande, J. D. (2011). Polymerase Chain Reaction : Methods , Principles and Application. International Journal of Biomedical Research, 2(1), 81–97.

Maftuchah, Winaya, A., & Zainudin, A. (2014). Tekhnik Dasar Analisis Biologi Molekuler. In A. Ikhwan (Ed.), Depublish (1st ed.). Depublish.

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 Science, 51(2), 143–148. https://doi.org/10.1016/S0309-1740(98)00112-0

Maulani, T. R., Susilo, H., Indriati, M., & Suhaemi, A. (2020). Deteksi Cemaran DNA Babi Dengan RT-PCR pada Sosis Tanpa Logo Halal di Kabupaten Pandeglang. Gorontalo Agriculture Technology Journal, 3(2), 72–80.

Nakanishi, H., Yoneyama, K., Hara, M., Takada, A., & Saito, K. (2021). Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing. Scientific Reports, 1–9. https://doi.org/10.1038/s41598-021-85803-4

Park, Y. H., Uzzaman, R., Park, J., & Kim, S. (2013). Detection of Meat Origin ( Species ) Using Polymerase Chain Reaction Detection of Meat Origin ( Species ) Using Polymerase Chain Reaction. July 2014. https://doi.org/10.5851/kosfa.2013.33.6.696

Perestam, A., Fujisaki, K., Nava, O., & Hellberg, R. (2017). Comparison of real-time PCR and ELISA-based methods for the detection of beef and pork in processed meat products. Food Control, 71, 346–352.

Popping, B., Diaz-Amigo, C., & Hoenicke, K. (2010). Chemists, Mollecular Biological and Immunological Techniques and Application for Food. John Wiley and Sons, Inc.

Prabawati, S. Y., & Fajriati, I. (2018). Analisis Lemak Sapi dan Lemak Babi Menggunakan Gas Chromatography (GC) dan Fourier Transform Infra Red Spectroscophy Second Derivative (FTIR-2D) untuk Autentifikasi Halal. Indonesia Journal of Halal, 1(2), 89. https://doi.org/10.14710/halal.v1i2.4119

Purnomo, N., Ramadhanty, D., Mansur, M., Nur, M., & Dagong, M. I. A. (2021). Kualitas DNA Hasil Purifikasi dari Sosis Sapi Sebagai Bahan Autentikasi Halal Berbasis Marker Genetik (DNA Quality of Purification from Beef Sausage as Halal Authentication Material Based on Genetic Marker). Jurnal Ilmu Dan Teknologi Peternakan, 9(1), 44–50. https://journal.unhas.ac.id/index.php/peternakan/article/view/11633/7219

Raharjo, T. J., Cahyaningtyas, W., & Pranowo, D. (2012). Validation Of PCR-RFLP Testing Method to Detect Porcine Contamination in Chicken Nugget. Indonesian Journal of Chemistry, 12(3), 302–307.

Sambrook, J., Fritsch, F., & T. Miniatis. (1989). Molecular Cloning Laboratory Manual. New York (US): Cold Spring Harbor Laboratory Pr. (3rd ed). Cold Spring Harbor Laboratory Press, Cold Spring Harbor.

Santoso, I., Mustaniroh, S. A., & Pranowo, D. (2018). Keakraban Produk dan Minat Beli Frozen Food: Peran Pengetahuan Produk, Kemasan, dan Lingkungan Sosial Product Familiarity and Purchase Intention of Frozen Food : The Role of Product Knowledge , Packaging , and Social Environment. 11(2), 133–144. https://doi.org/10.24156/jikk.2018.11.2.133

Seipp, M. T., Herrmann, M., & Wittwer, C. T. (2010). Automated DNA Extraction , Quantification , Dilution , and PCR Preparation for Genotyping by High-Resolution Melting. Journal of Biomolecular Techniques, 21(4), 163–166.

Sugiana, F. A., Widyowa, H., Warisman, M. A., Suryani, & Desriani. (2018). Low cost and comprehensive pork detection in processed food products with a different food matrix. Indonesian Journal of Biotechnology, 23(1), 21–27. https://doi.org/10.22146/ijbiotech.32372

Syukriya, A. J., & Faridah, H. D. (2019). Kajian Ilmiah dan Teknologi Sebab Larangan Suatu Makanan Dalam Syariat Islam. Journal of Halal Product and Research, 2(1), 47–48. https://e-journal.unair.ac.id/JHPR/article/download/13543/7598

Thangsunan, P., Temisak, S., Morris, P., Rios-solis, L., & Suree, N. (2021). Combination of Loop-Mediated Isothermal Amplification and AuNP-Oligoprobe Colourimetric Assay for Pork Authentication in Processed Meat Products. Food Analytical Methods, 14, 568–580. https://doi.org/https://doi.org/10.1007/s12161-020-01901-5

Zulaekah, S., & Kusumawati, Y. (2015). Halal dan haram makanan dalam Islam. SUHUF, 17(01), 25–35.




DOI: https://doi.org/10.17969/agripet.v23i2.26595

Article Metrics

Abstract view : 0 times
PDF - 0 times

Refbacks

  • There are currently no refbacks.


Copyright (c) 2023 Jurnal Agripet

License URL: http://www.jurnal.usk.ac.id/agripet/about/submissions#copyrightNotice


Creative Commons LicenseISSN: 1411-4623E-ISSN: 2460-4534
Copyright© 2000-2024 | ISSN: 1411-4623 | EISSN: 2460-4534 
Jurnal Agripet is licensed under a Creative Commons Attribution 4.0 International License.

 

Published by: 
Animal Husbandry DepartmentThe Faculty of Agriculture, Universitas Syiah Kuala 
associated with Animal Scientist's Society of Indonesia (HILPI)
Jl. Tgk. Hasan Krueng Kalee No. 3, Kopelma Darussalam,
Banda Aceh, 23111, Indonesia.
Phone: +62-81383736633
Email: jurnalagripet@usk.ac.id


Online Submissions & Guidelines Editorial Policies | Contact Statistics Indexing | Citations