Development And Application Of HRM For Detecting Porcine Stress Syndrome

Purpose:Hal is a major gene for porcine stress syndrome(PSS). PSS can influence the quality of meat, and individuals in transportation, drive or even breeding and other stress factors will burst malignant hyperthermia deaths, lead to a great loss to the porcine industry. And white muscle meat appears after abatage, reduce the meat quality. Therefore, it is extremely urgent to establish a simple and quick detection method of Hal.Method:According to the PSS RYRl gene sequences and mutational sites in GenBank,the two standard plasmids of C/C and T/T were synthesized, the genotype C/T also obtained after mixing the genotype C/C and T/T. Then two kinds of High Resolution Melting (HRM) curve anaysis:Small amplicon method and Unlabeled probe method, were established on the basis of these standard plasmids.use the two methods to detect the three hundred samples of porcine. In order to verification the accuracy of those two methods,those samples were detected by PCR-RFLP.Result: â‘ Three standard plasmids were successfully constructed. â‘¡Small amplicon method were successfully constructed and the detect result showed that those samples were no carriers of the Hal gene. â‘¢Unlabeled probe method were successfully constructed and the detect result showed that those samples were no carriers of the Hal gene. â‘£The detect result of PCR-RFLP showed that those samples were no carriers of the Hal gene.Conclusion:The results of the three detection methods are the same,so it can be used in the detection of Hal.