Tetraplex PCR assay involving double gene-sites discriminates beef and buffalo in Malaysian meat curry and burger products

M A Motalib Hossain; Md Eaqub Ali; Sharifah Bee Abd Hamid; S M Azad Hossain; Asing; Nina Naquiah Ahmad Nizar; Mohammad Nasir Uddin; Lokman Ali; Md Asaduzzaman; Md Jahurul Haque Akanda

doi:10.1016/j.foodchem.2016.12.062

Tetraplex PCR assay involving double gene-sites discriminates beef and buffalo in Malaysian meat curry and burger products

Food Chem. 2017 Jun 1:224:97-104. doi: 10.1016/j.foodchem.2016.12.062. Epub 2016 Dec 22.

Affiliations

¹ Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur 50603, Malaysia.
² Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur 50603, Malaysia; Centre for Research in Biotechnology for Agriculture (CEBAR), University of Malaya, Kuala Lumpur 50603, Malaysia. Electronic address: eaqubali@gmail.com.
³ Institute of Biological Sciences, University of Malaya, Kuala Lumpur 50603, Malaysia.
⁴ Department of Marine Bioresources Science, Faculty of Fisheries, Chittagong Veterinary and Animal Sciences University, Chittagong 4225, Bangladesh.
⁵ Faculty of Food Science and Nutrition, University Malaysia Sabah, Kota Kinabalu, Sabah 88400, Malaysia.

PMID: 28159299
DOI: 10.1016/j.foodchem.2016.12.062

Abstract

Replacement of beef by buffalo and vice versa is frequent in global markets, but their authentication is challenging in processed foods due to the fragmentation of most biomarkers including DNA. The shortening of target sequences through use of two target sites might ameliorate assay reliability because it is highly unlikely that both targets will be lost during food processing. For the first time, we report a tetraplex polymerase chain reaction (PCR) assay targeting two different DNA regions in beef (106 and 120-bp) and buffalo (90 and 138-bp) mitochondrial genes to discriminate beef and buffalo in processed foods. All targets were stable under boiling, autoclaving and microwave cooking conditions. A survey in Malaysian markets revealed 71% beef curries contained buffalo but there was no buffalo in beef burgers. The assay detected down to 0.01ng DNA and 1% meat in admixed and burger products.

Keywords: Biomarkers; Double gene targets; Target stability.

MeSH terms

Animals
Buffaloes
Cattle
Cooking / methods*
Cooking / standards
DNA / analysis
DNA / genetics
Food Contamination / analysis*
Food Handling / methods*
Food Handling / standards
Food Labeling / methods
Food Labeling / standards
Meat Products / analysis*
Meat Products / standards
Polymerase Chain Reaction / methods*
Polymerase Chain Reaction / standards
Red Meat / analysis*
Red Meat / standards
Reproducibility of Results

Substances

DNA