DETEKSI DAN DIFERENSIASI CAMPURAN DAGING BABI DAN SAPI DALAM PRODUK OLAHAN DAGING DENGAN METODE HIGH RESOLUTION MELT ANALYSIS (HRMA)

WINAHYU ESTRI, drh. Sidna Artanto, M. Biotech.

2014 | Skripsi | KEDOKTERAN HEWAN

Semakin maraknya modus pencampuran daging sapi dengan daging babi menyebabkan lembaga pengkajian makanan seperti Lembaga Pengkajian Pangan Obat-obatan dan Kosmetika Majelis Ulama Indonesia (LPPOM MUI) gencar dalam mengupayakan pemberantasan peredaran produk olahan yang telah tercemar daging babi. Beberapa metode deteksi seperti, Enzym Linked Immunosorbent Assay (ELISA), Fourier Transform Infrared Spectofotometry (FTIR), Polymerase Chain Reaction (PCR), dan Real-Time Polymerase Chain Reaction (Real-Time PCR) dinilai masih memiliki beberapa kekurangan. High Resolution Melt Analysis (HRMA) merupakan metode analisis DNA dengan real-time PCR yang mudah dilakukan karena kurva melting temperature akan langsung terbentuk setelah proses amplifikasi selesai dilakukan. Kurva Tm yang terbentuk didasari peningkatan pendaran warna yang dihasilkan oleh zat warna yang berpendar apabila terdapat untai ganda DNA. Pendaran warna pada proses ini dimonitori oleh pewarna evagreen dalam HRMA kit. Dalam penelitian ini digunakan sampel daging sapi dan babi murni serta sampel bakso yang diisolasi DNAnya. Sampel DNA diamplifikasi dengan metode HRMA menggunakan real-time PCR. Kurva melting temperature yang dihasilkan diidentifikasi berdasarkan melting temperature (Tm) masing-masing produk PCR. Kurva tersebut dianalisis dengan menggunakan software Bio-Rad CFX Manager 3.0. Kurva hasil penelitian menunjukkan adanya perbedaan signifikan melting temperature sampel (daging sapi, babi, bakso) pada kurva melting temperature yang terbentuk. Melting temperature sampel babi lebih tinggi dibandingkan dengan sampel sapi, dan melting temperature sampel bakso berada di antara kedua sampel tersebut. Hal tersebut menunjukkan bahwa sampel bakso mengandung daging babi dan daging sapi di dalamnya. Berdasarkan hasil tersebut, HRMA terbukti sebagai metode yang dapat digunakan untuk mendeteksi campuran daging babi dan sapi serta membedakan jenis kedua daging tersebut.

An increasingly widespread act of mixing beef meat with pork has caused several food assessment institutions such as Lembaga Pengkajian Pangan Obatobatan dan Kosmetika Majelis Ulama Indonesia (LPPOM MUI) intensified their effort in eradicating processed foods that are contaminated by pork meat. Several detection methods such as Enzym Linked Immunosorbent Assay (ELISA), Fourier Transform Infrared Spectofotometry (FTIR), Polymerase Chain Reaction (PCR), dan Real- Time Polymerase Chain Reaction (Real-Time PCR) are deemed to be flawed and somewhat lacking. High Resolution Melt Analysis (HRMA) is a method that could be applied to analyse DNA with real-time PCR that the melting curves will be immediately formed based on fluorescence produced by the dyes when there is a double-stranded DNA. Fluoresence dyes in that process is evagreen. This research uses pure beef and pork meat as well as meatball samples to acquire pure DNA results. The sample DNA is then amplified with HRMA method using real-Time PCR. The melting curve were identified based on the melting temperature (Tm) of each PCR product/amplicon. The melting curve were analysed using Bio-Rad CFX Manager 3.0. The result of the research has shown significant differences between the samples (beef, pork, and meatball) on the melting curve that was formed. Pork has a higher melting temperature then beef, and meatball sample has melting temperature in between pork and beef. It was shown that the meatballs sample containing pork and beef in it. Based on that result, HRMA is proven as a method that can be used to identify an adulteration in meat and differentiate the types of meat.

Kata Kunci : High Resolution Melt Analysis (HRMA), daging babi, daging sapi, DNA