Rapid detection of pork using alkaline lysis- Loop Mediated Isothermal Amplification (AL-LAMP) technique. (April 2020) - Girish, P.S.
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. Highlights: Manuscript reports a rapid technique for the species identification of pork. Technique uses Alkaline lysis method for the extraction of DNA. Target DNA was amplified using novel pork specific LAMP primers. Results can be visualized by colour changes after the addition of SYBR Green I dye. AL-LAMP assay has potential to be used at field level for pork authentication.