A novel duplex SYBR Green real-time PCR with melting curve analysis method for beef adulteration detection.

An efficient and reliable duplex SYBR Green real-time quantitative PCR (qPCR) method for beef products adulteration detection was developed based on bovine specific and vertebrate universal primers. By analyzing the numbers, positions (Tm value) of melting curve peaks of the duplex PCR products, we simultaneously identified bovine and preliminary screened non-bovine in samples, and also semi-quantified the bovine percentage according to the area ratios of peaks. All of these were necessary for adulteration determination.

A rapid and reliable multiplex PCR assay for simultaneous detection of fourteen animal species in two tubes.

A simple and rapid method for animal species identification to prevent food adulteration based on mitochondrial DNA using two independent multiplex polymerase chain reactions (PCRs) and microchip electrophoresis was developed. This method was designed to identify fourteen domestic animals (Group I: cattle, donkey, dog, fox, raccoon-dog, deer and horse; Group II: pig, sheep, goat, chicken, duck, cat and mouse) simultaneously using ten pairs of primers and three of which were degenerate primers. Sequences for species-specific primers were generated based on mitochondrial genes, including 12S rRNA, 16S rRNA, ND2 and CO I.