啮齿类螺杆菌SYBR Green Ⅱ荧光定量PCR检测方法的建立

Establishment of SYBR Green Ⅱ Based Real-time PCR Method for Detection of Helicobacter Rodentium

目的建立一种啮齿类螺杆菌(helicobacter rodentium)的快速检测方法。方法根据啮齿类螺杆菌16SrRNA的基因序列设计了1对特异性引物,建立了啮齿类螺杆菌SYBR GreenⅡ荧光定量PCR检测方法。结果在10~2拷贝/μL～10~7拷贝/μL浓度范围中标准曲线循环阈值与模板浓度呈良好的线性关系。该方法特异性良好,仅对啮齿类螺杆菌产生特异性扩增,且灵敏度可达到30拷贝/μL,组内和组间的变异系数均小于2%,具有良好的重复性。结论建立了SYBRGreenⅡ荧光定量PCR检测啮齿类罗杆菌的一种方法 。

Objective To establish a rapid detection method for helicobacter rodentium. Method A pair of specific primers was designed according to the gene sequence of Helicobacter rodentium 16S rRNA. The method of SYBR Green Ⅱ real-time PCR was established. Results The standard curve cycle threshold had a good linear relationship with the template concentration in the range of 102 copies/μL to 107 copies/μL concentration. This method has good sensitivity, only specific amplification of Helicobacter rodentium, could detect 30 copies/μL, the coefficient of variation （CV） was less than 2% within and between groups with good reproducibility. Conclusion The method of SYBR Green II real- time PCR was established for the identification of Helicobacter rodentium.