Ultra-fast DNA-based multiplex convection PCR method for meat species identification with possible on-site applications

- Ultra-fast multiplex meat identification method that detected meat species in 24 min was developed.
- As low as 1 pg of genomic DNA of meat species could be detected.
- The method could detect as little as 1% adulteration.
- The method could be used identification of raw and thermal processed meats.
- Sampling, DNA isolation, convection PCR and electrophoresis could be performed in less than 60 min.

The aim of this study was to develop an ultra-fast molecular detection method for meat identification using convection Palm polymerase chain reaction (PCR). The mitochondrial cytochrome b (Cyt b) gene was used as a target gene. Amplicon size was designed to be different for beef, lamb, and pork. When these primer sets were used, each species-specific set specifically detected the target meat species in singleplex and multiplex modes in a 24 min PCR run. The detection limit was 1 pg of DNA for each meat species. The convection PCR method could detect as low as 1% of meat adulteration. The stability of the assay was confirmed using thermal processed meats. We also showed that direct PCR can be successfully performed with mixed meats and food samples. These results suggest that the developed assay may be useful in the authentication of meats and meat products in laboratory and rapid on-site applications.