Abstract
A highly specific single step polymerase chain reaction (PCR) is described for the detection of pig (Sus domesticus) meat. A PCR assay was successfully optimized for amplification of 629 and 322-bp DNA fragment extracted from pig meat using designed species-specific primer pairs based on mitochondrial D-loop and 12Sribosomal ribonucleic acid (rRNA) gene, respectively. The optimized PCR assay was subsequently validated for its specificity with deoxyribonucleic acid (DNA) extracted from cattle, buffalo, sheep, goat and pig. PCR amplification of target DNA with pig-specific primers was repeated 15 times, with consistent results observed. The specificity of pig-specific PCR provides a valuable tool for identification of pig meat and to avoid its fraudulent substitution and adulteration. Key words: Pig meat, adulteration, polymerase chain reaction (PCR), mitochondrialD-Loop, 12S ribosomal ribonucleic acid (rRNA) gene
Highlights
The adulteration/substitution of meat has always been a concern for various reasons such as public health, religious factors, wholesomeness and unhealthy competition in meat market (Arslan et al, 2006)
A polymerase chain reaction (PCR) assay was successfully optimized for amplification of 629 and 322-bp deoxyribonucleic acid (DNA) fragment extracted from pig meat using designed species-specific primer pairs based on mitochondrial D-loop and 12S ribosomal ribonucleic acid gene, respectively
The optimized PCR assay was subsequently validated for its specificity with deoxyribonucleic acid (DNA) extracted from cattle, buffalo, sheep, goat and pig
Summary
The adulteration/substitution of meat has always been a concern for various reasons such as public health, religious factors, wholesomeness and unhealthy competition in meat market (Arslan et al, 2006). Species-specific PCR assay was found to be rapid and cost effective for identification of meat species due to specific detection of target sequence without the need of further sequencing or digestion of the PCR products with restriction enzymes (Rodriguez et al, 2004) and successfully used for identification of various species of meat (Ilhak and Arslan, 2007; Martin et al, 2007; Frezza et al, 2008; Mane et al, 2007) These PCR assays targets genomic as well as mitochondrial DNA for the purpose of meat species identification. We describe a species specific PCR assay which holds a unique advantage over other DNA based methods in terms of specificity, sensitivity, robustness and rapidity Keeping these considerations in view, the present work was designed to develop highly specific PCR using newly developed primers for identification of pig meat in raw meat samples
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