Tetraplex PCR assay involving double gene-sites discriminates beef and buffalo in Malaysian meat curry and burger products.
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.
Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur 50603, Malaysia.
M A Motalib
MA
Hossain
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.
Md Eaqub
ME
Ali
Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur 50603, Malaysia.
Sharifah Bee Abd
SB
Hamid
Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur 50603, Malaysia.
S M Azad
SM
Hossain
Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur 50603, Malaysia.
Asing
Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur 50603, Malaysia.
Nina Naquiah Ahmad
NN
Nizar
Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur 50603, Malaysia.
Mohammad Nasir
MN
Uddin
Institute of Biological Sciences, University of Malaya, Kuala Lumpur 50603, Malaysia.
Lokman
L
Ali
Department of Marine Bioresources Science, Faculty of Fisheries, Chittagong Veterinary and Animal Sciences University, Chittagong 4225, Bangladesh.
Md
M
Asaduzzaman
Faculty of Food Science and Nutrition, University Malaysia Sabah, Kota Kinabalu, Sabah 88400, Malaysia.
Md Jahurul Haque
MJ
Akanda
D000818:Animals; D002020:Buffaloes; D002417:Cattle; D003296:Cooking / Q000379:methods* / Q000592:standards; D004247:DNA / Q000032:analysis / Q000235:genetics; D005506:Food Contamination / Q000032:analysis*; D005511:Food Handling / Q000379:methods* / Q000592:standards; D005515:Food Labeling / Q000379:methods / Q000592:standards; D008461:Meat Products / Q000032:analysis* / Q000592:standards; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D000069466:Red Meat / Q000032:analysis* / Q000592:standards; D015203:Reproducibility of Results
D004247:DNA
Biomarkers; Double gene targets; Target stability