Forgery Detection Beef with Mice Meat ( Mus musculus ) in Meatballs Using Real-Time Polymerase Chain Reaction (Real-Time PCR) Primer Specific for a Target Mitochondrial DNA ND-1 Gene

Tri Joko Raharjo; Gilang Aji Pratama; Irma Nuryanti; Rarastoeti Pratiwi

doi:10.22146/ijc.27542

Forgery Detection Beef with Mice Meat (Mus musculus) in Meatballs Using Real-Time Polymerase Chain Reaction (Real-Time PCR) Primer Specific for a Target Mitochondrial DNA ND-1 Gene

https://doi.org/10.22146/ijc.27542

Tri Joko Raharjo^(1*), Gilang Aji Pratama⁽²⁾, Irma Nuryanti⁽³⁾, Rarastoeti Pratiwi⁽⁴⁾

(1) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281, Indonesia
(2) Biotechnology Study Program, Graduated School, Universitas Gadjah Mada, Yogyakarta
(3) Dexa Laboratories of Biomolecular Sciences, Cikarang, Bekasi 17530, West Java, Indonesia
(4) Faculty of Biology, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281, Indonesia
(*) Corresponding Author

Abstract

The expensive beef have encouraged counterfeiting beef on processed food products such as meatballs. Mice meat is frequently reported used for adulteration of beef. The accurate method is needed to ensure the supervision of food safety. This study reports the use of DNA testing to detect the presence of mice meat in meatballs with real-time PCR primer specific. PCR primers designed based on the ND-1 gene of mice mitochondrial DNA with the sequence are 5’-CGGCATCCTACAACCATTTGC-3’ and 5’-CGGCTCGTAAAGCTCCGAA-3’, respectively, target 294 bp DNA fragment. The real-time PCR can specifically detect the presence of the mice meat in a meatball with no detection the presence of beef, mutton, chicken, pork, and horsemeat. The method showed good precision shown by the CV of repeatability test at 2%, much lower than the requirement of < 25%. Real-time PCR was able to deliver positive results for as low as 0.5 ng DNA template, equivalent to 0.08 copies of genome DNA of mice equal to 80–150 copies of mtDNA. By using standard phenol-chloroform DNA isolation technique, this method is able to detect contamination of mice meat in meatball up to 1%. Three commercial meatballs confirmed contaminated by mice meat using the method.

Keywords

mice; meatball; real-time PCR; ND-1; mitochondrial DNA

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DOI: https://doi.org/10.22146/ijc.27542