基于重组酶辅助扩增技术的绵羊与山羊源性成分快速鉴别OA

In order to solve the problem of meat adulteration caused by the difficulty in distinguish between sheep and goat meat due to the loss of phenotypic characteristics after processing,a dual real-time fluorescence method based on recombinase-aided amplification(RAA)was developed for the simultaneous detection of sheep-and goat-derived components.Primers and probes targeting the conserved region of the Cytb gene of sheep and goat mitochondrial DNA were designed and screened.The reaction temperature was optimized.The results showed that the combination of the goat-specific primer Capra-F2/R2 and the sheep-specific primer Ovis-F2/R1 had the best amplification performance.The method could be completed at 39℃for 30 min without complex thermal cycling steps.The limits of detection for sheep-and goat-derived components in the mixed system of goat and sheep meat reached 0.5%(m/m),and there was no cross-reaction with 16 common meat species such as beef,pork,and duck meat.This method could accurately identify sheep-and goat-derived components in actual samples,and the results were completely consistent with those of real-time fluorescence quantitative polymerase chain reaction(real-time PCR),with an accuracy rate of 100%.In summary,the dual real-time RAA method is rapid,sensitive,highly specific,and less dependent on equipment.It is suitable for the rapid identification of sheep-and goat-derived components,providing technical support for the authenticity identification of meat.