摘要
目的建立高效特异的巴尔通体实时荧光定量PCR检测方法,并应用于实验用猫的微生物检测工作中。方法针对NCBI公布的巴尔通体序列设计特异引物和Taq Man探针,使用分子生物学方法制备质粒标准品,建立巴尔通体实时荧光定量PCR方法;对该方法的线性、敏感性、特异性及稳定性进行测定;并使用该方法对142个猫样品进行检测。结果成功建立巴尔通体实时荧光定量PCR方法;该方法线性范围为1.0×101copies/μL^1.0×109copies/μL,相关系数为0.998,检测极限达10 copies/μL;特异性结果显示所建方法具有良好的特异性,并在142份实验用猫样品中检测出阳性样品6份。结论实时荧光定量PCR方法可用于实验用猫巴尔通体的检测工作中。
Objective To establish an effective and specific real-time PCR assay for Bartonella detection, and use the assay in experimental cat. Method According Bartonella RIBC gene fragments, a pair of specific primers and TaqMan probe were designed and synthesized. Bartonella DNA standards were prepared using some molecular biological method . Then the linearity, sensitivity, specificity and stability of the method were measured. At last, the method was used for testing blood samples from 142 cats. Result We established successfully real-time PCR method for Bartonella detection. The linear range was 1.0 × 10^1 - 1.0 × 10^9 copies/μL, the lowest sensitivity was 10 copies/μL, the coefficient of variation （CV） was 0. 998. There was no false positive detection from other bacterial strains. 6 positive reactions were detected in the blood samples of 142 cats. ConcLusion The method is able to be used to detect and quantify the Bartonella in experimental cats.
作者
冯育芳
邢进
王吉
付瑞
岳秉飞
FENG Yufang;XING Jin;WANG Ji;FU Rui;YUE Bingfei(Institute for Laboratory Animal Resources,National Institutes for Food and Drug Control,Beijing 100050,China)
出处
《实验动物科学》
2018年第4期56-60,共5页
Laboratory Animal Science
基金
北京市科技计划(No.D171100002717001)
