实时荧光聚合酶链反应检测空肠弯曲菌

Detection of Campylobacter jejuni by real-time fluorescent PCR

目的探索一种快速、敏感、特异的空肠弯曲菌检验方法。方法根据GenBank公布的空肠弯曲菌基因序列,在其保守区域设计特异性引物与探针以建立基于TaqM an探针的实时荧光聚合酶链反应(PCR)方法,并对方法的特异性、敏感性和重复性进行研究。结果该方法对空肠弯曲菌可产生特异扩增信号,对其他非空肠弯曲菌菌属无响应,其检测敏感性可达10 cfu/mL。反应体系具有很高的稳定性。结论实时荧光PCR可快速、特异、灵敏地检测空肠弯曲菌,具有较强的实际应用价值。

Objective To explore a rapid, sensitivety and specific method for examining Campylobacterjejuni （C. jejuni）. Methods According to C. jejuni sequences from Ganbank, the specific primers and probe were designed on the basis of conserved gene region and a TaqMan real-time fluorescent polymerase chain reaction （PCR） was established. The specificity, sensitivity and repeatability of the assay were also studied. Results C. jejuni was amplified by real - time fluorescent PCR successfully while all non-C, jejuni bacterial species did not generate any signal. The assay had high sensitivity and repeatability, and the low detecting limit was 10 cfu/mL. Conclusions The real-time fluorescent PCR is a rapid, specific and sensitive method for quantifying C. jejuni, thus it has high practical value of application.