鼠巴贝斯虫TaqMan探针荧光定量PCR方法的建立及应用

Development and application of a TaqMan real-time PCR method for the detection of B.microti

目的建立一种用于检测鼠巴贝斯虫的TaqMan探针荧光定量PCR方法。方法针对鼠巴贝斯虫18sRNA基因序列,设计特异性引物和探针,建立鼠巴贝斯虫的TaqMan探针荧光定量PCR反应体系,并对该方法的灵敏度和特异性进行验证。结果本方法对鼠巴贝斯虫的检测灵敏度较高,最低检出限为1.34×10-7ng/μl;具有较高的特异性,与钩端螺旋体、莱姆病螺旋体、巴尔通体、Q热、土拉菌等病原均无交叉反应,反应体系稳定。结论本研究建立的TaqMan探针荧光定量PCR方法特异、灵敏,可用于鼠巴贝斯虫的实验室快速检测及现场监测使用。

Objective To develop a TaqMan real-time PCR method to detect B.microti. Methods Specific primers and probes were designed according to the sequences of 18 sRNA gene of B.microti, to set up a TaqMan real-time PCR method to detect B.microti. The sensitivity and specificity were also analyzed. Results The assay can detect B.microti with high specificity and the lowest detection limitation was 1.34 ×10-7ng/μl. Conclusion The assay, which is high sensitivity and specificity, can be used to detect B.microti in the laboratory.