A recent study (here – open access) reported the development and validation of a duplex real-time polymerase chain reaction (qPCR) assay for simultaneous identification of sheep and goat meat. Amplicons were detected using high-resolution melt analysis (HRMA). Species-specific amplicons with melting temperature of 78 °C for goat and 80 °C for sheep (with associated standard errors) were identified. The assay was optimized for basic PCR parameters and the HRMA detection was standardized based on melt curve shape, colour and inflexion points. The absolute limit of detection (LODabs) and limit of quantification (LOQ) of the assay were found to be 0.39 ng and 0.78 ng of target DNA respectively. A sensitivity of 0.1% (w/w of meat) was obtained for both sheep and goat meat. The linearity and efficiency were evaluated in both simplex and duplex assays. The authors concluded that this closed-tube qPCR-HRMA method can be a rapid and sensitive detection technique for meat speciation.
Photo by Ali Burhan on Unsplash
Comments