Abstract:

In this study, a target-enriched multiplex PCR (Tem-PCR) assay for simultaneous detection of Salmonella,
Staphylococcus aureus and Shigella was developed. Based on the S. aureus femA gene, the Salmonella invA gene and
the Shigella ipaH gene, three pairs of specific primers together with a universal primer were designed. Following the
optimization of reaction conditions, the Tem-PCR assay was successfully established. Results showed that the Tem-
PCR assay was highly specific to the target bacteria with a sensitivity of 1.1 × 103 CFU/mL, 2 × 103 CFU/mL and
1.2 × 103 CFU/mL for Salmonella, S. aureus and Shigella, respectively. The Tem-PCR assay can effectively improve the
balance of amplification efficiencies of the primers employed in traditional multiplex PCR.

Key words: Tem-PCR assay, Salmonella, Staphylococcus aureus, Shigella