应用实时荧光PCR技术检测空肠弯曲菌

Application of real-time fluorogenic PCR assay for the detection of Campylobacter jejuni

目的应用实时荧光PCR技术建立一种特异、敏感的空肠弯曲菌(Campylobacter jejuni)检测方法。方法通过对空肠弯曲菌基因组的保守序列进行分析来设计引物及Taqman探针来实现对空肠弯曲菌的检测。结果该方法具有较强的灵敏性及特异性能够对空肠弯曲菌进行有效的检测,其对空肠弯曲菌增菌液的最低检出限为5CFU/ml而对实际样品的检测限为10CFU/g。结论该方法无需增菌培养过程能够直接对样品进行检验,较常规的生化检测方法省时省力,具有较强的实际应用价值。

The real-time fluorogenic PCR assay was used for the detection of Campylobacter jejuni by using the primers designed from the conserved genomic sequence of C. jejuni and Taqman probe. This method was proved to be sensitive and specific to detect C. jejuni efficiently. It was demonstrated that the lowest detection limit of this method for bacterial suspensions of C. jejuni was 5 CFU/ml, while that for clinical specimens was 10 CFU/g. It is evident that this method can be directly used to detect C.jejuni from clinical specimens without cultivation of the organisms, thus have more practical value of application.