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当前位置: 首页 > 文章 > 布氏杆菌微滴数字PCR方法的建立 畜牧与兽医 2022,54 (2) 97-101
Position: Home > Articles > Development of droplet digital PCR for the detection of Brucella strains Animal Husbandry & Veterinary Medicine 2022,54 (2) 97-101

布氏杆菌微滴数字PCR方法的建立

作  者：
董浩;原霖;刘洋;吴同垒;陈亚娜;徐阳;王传彬;梁春南
单  位：
河北科技师范学院;中国动物疫病预防控制中心;中国食品药品检定研究院
关键词：
布氏杆菌;微滴数字PCR;实时荧光PCR
摘  要：
为建立一种布氏杆菌微滴数字PCR qPCR方法,对布氏杆菌病的定量诊断提供技术支持,在实时荧光PCR(qPCR)检测方法(T/CVMA 20—2020)的基础上,建立了布氏杆菌微滴数字PCR方法,并对方法的反应条件进行了优化,同时对其敏感性、特异性、重复性进行了评估。结果显示:本方法的最低检测下限为2.6 copies/μL,未发现与常见的5种细菌存在交叉反应,重复性检测的变异系数小于5%;采用该微滴数字PCR和实时荧光PCR检测方法分别对76份流产牛的阴道拭子进行检测,显示该微滴数字PCR比qPCR可以多检出3份阳性样品。本研究建立的微滴数字PCR方法敏感性高、特异性强、重复性好,适用于临床样品的布氏杆菌核酸检测。
译  名：
Development of droplet digital PCR for the detection of Brucella strains
作  者：
DONG Hao;YUAN Lin;LIU Yang;WU Tonglei;CHEN Yana;XU Yang;WANG Chuanbin;LIANG Chunnan;National Institutes for Food and Drug Control;China Animal Disease Control Center;Hebei Normal University of Science and Technology;
关键词：
Brucella spp.;;droplet digital PCR;;real-time PCR
摘  要：
A high-sensitivity droplet digital PCR method was to be established here for quantitative detection of Brucellosis. According to the real-time PCR method for the detection of Brucella spp. in animals(T/CVMA 20—2020), a droplet digital PCR method for Brucella spp. detection was developed. The reaction system of the method was optimized. The limit of detection, specificity and repeatability of this method were evaluated, and results showed that its limit of detection was 2.6 copies/μL. No cross-reaction of the method with 5 common bacteria was observed, and the coefficient of variation of the sample repetition was less than 5%. 76 vaginal swabs collected from aborted cows were detected by the developed droplet digital PCR and qPCR method, respectively, and the results showed that three more positive samples were detected by droplet digital PCR than by qPCR. Therefore, the droplet digital PCR method established in this study had good sensitivity and specificity, and would be helpful in detection of the nucleic acid of Brucella spp. in clinical practice.

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