食品中铜绿假单胞菌实时荧光PCR检测的研究

摘要

A fluorescence real-time PCR method was developed to detect the Pseudomonas aernginosa, Species-specific primers and TaqManfluorescent probe were designed by Primer express 3.0 software according to the conserved sequence of Exotoxin Agene of Pseudomonas aernginosa. Different gradient concentrations of Pseudomonas aernginosa DNA and various pathogen DNA were amplified by fluorescent real-time PCR to con-firm the specificity and sensitivity of the developed method. Results showed that the amplification curves were expressed from Pseudomonas aernginosa by fluorescence real-time PCR.The sensitivity of fluorescence real-time PCR for Pseudomonas aernginosa was 1×103 CFU/mL. This method is valuable for research and application prospects.%根据铜绿假单胞菌外毒素A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,建立铜绿假单胞菌实时荧光PCR检测方法.通过对梯度含量的铜绿假单胞菌标准菌液样品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.试验结果表明,只有铜绿假单胞菌产生扩增曲线,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1×103 CFU/mL.该方法具有很好的研究价值和应用前景.