Vol. 12, Issue 9 (2023)

The present study aims at development of a novel Polymerase Spiral Reaction (PSR) assay to detect the presence of goat meat (chevon) in adulterated meat. Many PSR assays have been developed to identify bacteria, fungi and even viruses. But, PSR based assays have not yet been reported for confirming meat species, to the best of our knowledge. The DNA from meat samples were extracted employing the alkaline lysis (AL) technique. The extracted DNA were evaluated for its purity by spectrophotometer analysis. The samples had an Optical Density (OD) ratio of 1.6-1.8 between a wavelength of 260 and 280 nm. In this study, a pair of PSR chevon-specific primers were sequenced targeting goat specific mitochondrial DNA (mtDNA). The possibility of cross-amplification of the primers was ruled out by considering related species. The results were analysed by (1) Observation of visual colour change by the addition of SYBR Green I stain to the amplified product, (2) Measuring the intensity of colour developed and (3) Visualization of bands on agarose gel electrophoresis (2%). The specificity of the chevon-specific PSR was determined using DNA extracted from meat samples obtained from different meat species viz., buffalo, cattle, chicken, pig, sheep and ostrich. There was no cross amplification observed when the PSR assay was carried out with DNA of other species. The experiment was repeated six times and consistent results were obtained. Results obtained in the present study are reliable and can be used for field purposes which take about 90 min when combined with the alkaline lysis method of DNA extraction.