Rapid detection of pork using Alkaline lysis - Loop mediated isothermal amplification (AL-LAMP) technique

Abstract

We report a novel, rapid, economical and species-specific DNA-based assay for the authentication of pork. The technique specifically amplified porcine mitochondrial D loop region by combining Alkaline Lysis (AL) method of DNA extraction and Loop Mediated Isothermal Amplification (LAMP). Visual detection of the reaction was accomplished by color development in the reaction with the addition of SYBR Green I dye. Dependable amplification was possible in thermally processed meat samples heated up to 121 °C for 30 min. The assay was able to detect pork in beef up to the level of 0.1% admixture and limit of detection of DNA was at 0.5 ng/μL. Cross-amplification of related species like cattle, buffalo, sheep, goat and chicken was excluded by incorporating their DNA in the reaction assay. The novel approach (AL-LAMP technique) was found to be robust and handy, suitable even for resource compromised laboratories engaged in the food analysis.