Abstract:A helicase-dependent isothermal DNA amplification (HAD) method is used for rapid and accurate detection Staphyloccocus aureus in pork, beef, mutton and chicken. A pair of oligonucleotide primers exclusively amplified the nuc gene of Staphyloccocus aureus, thus, Staphyloccocus aureus could be directly detected in pork, beef, mutton and chicken. The final concentrations of UvrD helicase and T4 gp32 were optimized. The amplification product was detected by electrophoresis. The results showed that HAD method had high specificity in detection of Staphyloccocus aureus in meat, but the other bacteria could not been amplificated. The amplification product had the same 216 bp length as the designed gene fragment, and the detection limit was 101 CFU/g. The optimized concentrations of UvrD helicase and T4 gp32 were 0.1 μg and 5.0 μg, respectively. HDA method was faster and more sensitive for detection of Staphyloccocus aureus in pork, beef, mutton and chicken, and established the foundation for the rapid detection of Staphylococcus aureus in other foods.
