中国动物检疫Issue(8)：40-42,3.
新孢子虫荧光PCR检测方法的研究
Research of Real-time PCR for Detection of Neospora Caninum
摘要
Abstract
A pair of specific primers and probe were designed according to the published Nc-5 gene sequence of Neospora caninum (N.caninum)in GeneBank. After the optimization of Mg2+ concentration,primer concentration,probe concentration,dNTP concentration,Taq enzyme and anneal temperature,a real-time PCR method was established to detect N.caninum infections. The sensitivity of the method for NC-5 recombinant plasmid was 10 copies per reaction. The coefficient of variation(CV)for repeated detection of serial dilutions of recombinant plasmid was 0.50%~1.18%.50 cattle blood sample and 8 cattle embryo samples of miscarriages were detected by the method,with 5 blood samples and 1 fetus sample as positive. The results showed that the average positive ratio was 10.3%,higher than the common PCR(7%).The method had good specificity and reliability,it could be used to detect Neospora caninum.关键词
新孢子虫/Nc-5基因/荧光PCRKey words
Neospora caninum/NC-5 gene/Real-time PCR分类
农业科技引用本文复制引用
员丽娟,史茜,季新成,牛国辉..新孢子虫荧光PCR检测方法的研究[J].中国动物检疫,2012,(8):40-42,3.基金项目
国家质量监督检验检疫总局科研项目(资助编号:2009IK011,2011IK017) （资助编号:2009IK011,2011IK017）
