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

作　　者：陈倩[1] 王静[1] 侯咏 富英群 李明 呼满霞[3] 杨军 王旺[1] 郭天宇[1] 刘丽娟[1] 

机构地区：[1]中国检验检疫科学研究院卫生检疫研究所,北京100123 [2]黑龙江出入境检验检疫局 [3]绥芬河出入境检验检疫局 [4]逊克出入境检验检疫局

出　　处：《中国国境卫生检疫杂志》2014年第4期221-225,共5页Chinese Journal of Frontier Health and Quarantine

基　　金：质检公益项目(201310076;201310072);国际合作项目(2012DFA30540)和(CAIQ2012JK038)

摘　　要：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.

关 键 词：TaqMan荧光定量PCR 鼠巴贝斯虫 应用 

分 类 号：R382.9[医药卫生—医学寄生虫学]