肉制品中表皮葡萄球菌实时荧光PCR检测方法的研究

Study on real-time fluorescent PCR detection method of Staphylococcus epidermidis in meat products

根据表皮葡萄球菌16S基因片段序列,用Primer express 3.0设计一对特异性引物和一个TaqMan探针,建立表皮葡萄球菌实时荧光PCR检测方法。通过对梯度含量的表皮葡萄球菌标准菌液样品DNA和多种细菌的DNA进行实时荧光PCR检测,来检测其灵敏度和验证引物和探针的特异性。实验结果表明,只有表皮葡萄球菌产生扩增曲线,其他细菌无扩增,说明引物及TaqMan探针特异性较好,检测灵敏度为1×10~3CFU/mL。该方法具有很好的研究价值和应用前景。

According to the 16 S gene fragment sequence of Staphylococcus epidermidis,primer express 3.0 was used,and a pair of specific primers and a TaqMan probe were designed,and a real-time fluorescent PCR detection method for Staphylococcus epidermidis was established.Gradient content of Staphylococcus epidermidis standard bacterial liquid DNA and various bacterial DNA were detected by real-time fluorescent PCR,and the sensitivity was detected,and the specificity of the primers and probes was verified.The experimental results showed that only Staphylococcus epidermidis produced an amplification curve,and other bacteria did not amplify,it indicated that the primers and TaqMan probes had good specificity,and the detection sensitivity was 1×10~3 CFU/mL.This method had good research value and application prospects.