A novel multiplex-PCR assay to detect three non-halal meats contained in meatball using mitochondrial 12S rRNA gene

Muhammad Cahyadi¹^,^*, Tommy Wibowo¹, Ahmad Pramono¹, Zakaria Husein Abdurrahman²

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¹Department of Animal Science, Faculty of Agriculture, Sebelas Maret University, Surakarta 57126, Indonesia.

²Faculty of Animal Science, Boyolali University, Boyolali 57315, Indonesia.

^*Corresponding Author: Muhammad Cahyadi, Department of Animal Science, Faculty of Agriculture, Sebelas Maret University, Surakarta 57126, Indonesia. Phone: +62271-637-457. E-mail: mcahyadi@uns.ac.id.

© Copyright 2020 Korean Society for Food Science of Animal Resources. This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Received: Feb 27, 2020 ; Revised: May 11, 2020 ; Accepted: May 28, 2020

Published Online: Jun 05, 2020

Abstract

The objective of this study was to detect three non-halal meat products consisted of dog, pork, and rat species in beef meatball using novel multiplex-PCR with 12S rRNA gene as target sites. A total of 33 self-made meatballs were used, and they were grouped into eleven types of meatball based on meat species origin contained in the meatballs. Each type consisted of three meatballs. Extraction of genomic DNA from the meatballs was used as a DNA template for simplex-, duplex-, and multiplex-PCR processes. The result of simplex-PCR, duplex-PCR, and multiplex-PCR showed that the 12S rRNA primer gene successfully amplified DNA for each species bovine, dog, pig, and rat, which are respectively indicated by 155, 244, 357, and 491 bp of DNA bands. In addition, multiplex-PCR with 12S rRNA gene primers can be uniquely and accurately used for detection bovine, dog, pig, and rat species on beef meatball in one reaction.

Keywords: meatball; multiplex-PCR; non-halal; species; 12S rRNA