UJI SPESIFISITAS METODE DETEKSI CEMARAN DAGING TIKUS DALAM BAKSO SAPI MENGGUNAKAN TEKNIK POLYMERASE CHAIN REACTION (PCR)-PRIMER SPESIFIK BERDASARKAN URUTAN DNA MITOKONDRIA; SPECIFICITY TESTING OF DETECTION METHOD OF RAT MEAT CONTAMINATION IN BEEF MEATBALLS USING SPECIFIC PRIMERPOLYMERASE CHAIN REACTION (PCR) TECHNIQUE BASED ON MITOCHONDRIAL DNA SEQUENCE

Irma Nuryanti, Tri Joko Raharjo

2014 | Disertasi | PROGRAM STUDI S2 KIMIA

Specificity testing of detection method of rat meat contamination in beef meatballs using specific primer-polymerase chain reaction (PCR) technique based on mitochondrial DNA sequence have been carried out. This study aimed to create a specific primer derived from mitochondrial rat (Mus musculus castaneus) genome as the DNA marker for detection of rat meat in meatballs. The primer was designed using primer-BLAST software. Testing of primer performance was conducted by amplification of specific primer-PCR using DNA template of rat meatballs as positive control and beef meatballs as negative control. Specificity of the primer was also studied by comparing DNA template of rat to five other species (cattle, chicken, pig, horse and goat). Meatballs made of beef meat with addition of rat meat (1, 2, 5, 10, 25 and 50% (w/w)) were tested to obtained sensitivity of the primer. Primer ND1-P1 forward (5’-CGGCATCCTACAACCATTTGC-3’) and reverse (5’-CGGCTCGTAAAGCTCCGAA-3’) with amplification product size of 294 bp is selected from 9 primer candidates based on its performance. The specific fragment of ND1-P1 primer only amplify the target fragment in rat DNA but not in other species. Rat meat in meatballs can be detected at concentration as low as 5% (w/w). Therefore, the ND1 gene is potentially as specific marker for detection of rat meat in the meat products.

Kata Kunci : mitochondrial DNA, rat, specific primer-PCR, meat products