Screening of RYR1 genotypes in swine population by a rapid and sensitive method

Genetic polymorphism in the gene controlling the calcium release channel (ryanodine receptor, RYR1) of sarcoplasmic reticulum in skeletal muscle have been shown to affect meat quality in pigs, producing porcine stress syndrome (PSS) and malignant hyperthermia (MH). The MH condition produces pale, soft and exudative (PSE) meat resulting in reduced commercial value of pork. Therefore, monitoring swine population for RYR1 genetic variants is important in the subsequent process of breeding and improving meat quality in pigs. The aim of the present study was to establish and validate a rapid and sensitive molecular genotyping assay to detect C1843T mutations of the Sus scrofa RYR1 gene using quantitative PCR (qPCR) followed by high-resolution melting (HRM) analysis. The qPCR-HRM assay allowed the rapid and sensitive identification of RYR1 genotypes for discrimination of wild type and mutant alleles, so the carriers can be detected. The method was validated by analyzing a total of 195 DNA samples that were previously genotyped by PCR-RFLP method. The results demonstrate that qPCR-HRM method is a fast, simple and reliable assay for genotyping the C1843T (Arg615Cys) polymorphism of the RYR1 gene. The high frequency of RYR1 mutant allele suggested that implementing a routine testing system is necessary to gradually eradicate the MH condition.