应用实时荧光定量PCR方法检测实验用猫巴尔通体的感染  

Establishment and Application of Real time PCR Method for Bartonella Detection in Experimental Cat

作　　者：冯育芳[1] 邢进[1] 王吉[1] 付瑞[1] 岳秉飞[1] 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)

机构地区：[1]中国食品药品检定研究院实验动物资源研究所,北京100050

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)

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.

关 键 词：实验用猫 巴尔通体 实时荧光定量PCR 应用 

分 类 号：R332[医药卫生—人体生理学]