Development of qualitative and quantitative PCR analysis for meat adulteration from RNA samples

•We develop PCR-based methods for identification of meat adulteration from RNA samples.•Species-specific primers are designed from TnI, MRP and tropomodulin genes.•Long time storage at low temperature of meats is successfully differentiated.•Quantitative real-time PCR for meat adulteration is established from RNA samples. Total RNA samples were used to establish qualitative and quantitative PCR-based methods for assessing meat adulteration. The primers were designed based on the mRNA sequences of troponin I (TnI), mitochondrial ribosomal protein (MRP) and tropomodulin genes to distinguish chicken, pork, goat, beef and ostrich. There was no cross reaction between the primers, and the detection limit of the cDNA template was 0.01 and 20ng in simplex PCR and multiplex PCR, respectively. In the low temperature storage test, the detection limits of cDNA template with 10 and 1ng were determined at 4°C and −80°C. In quantitative assay, the precision of real-time PCR analysis expressed as a coefficient of variation (CV) ranged from 0.25% to 5.24% and the trueness, expressed as an error, ranged from 0.28% to 6.98% for adulteration. Thus, herein, we provided alternative tools for the assessment of meat adulteration using mRNA-based PCR methods.