绵羊血液中布氏杆菌DNA提取方法的比较研究  

Comparison of methods for extraction of Brucella DNA from sheep blood

作　　者：丁可莉 杨发龙[1] 张焕容[1] 郭莉[2] 阳爱国[2] 莫茜[2] 袁东波[2] 侯巍[2] DING Keli;YANG Falong;ZHANG Huanrong;GUO Li;YANG Aiguo;MO Xi;YUAN Dongpo;HOU Wei(College of Life Science and Technology,Southwest Minzu University,Chengdu 611130,China;Center for Animal Disease Prevention and Control of Sichuan Province,Chengdu 611130,China)

机构地区：[1]西南民族大学生命科学与技术学院,四川成都611130 [2]四川省动物疫病预防控制中心,四川成都611130

出　　处：《畜牧与兽医》2020年第3期78-82,共5页Animal Husbandry & Veterinary Medicine

基　　金：四川省科技厅项目《动物布氏杆菌病防控关键技术研究与应用》(2018JY0039);西南民族大学研究生创新科研项目(CX2019SZ110)。

摘　　要：本研究旨在比较5种DNA提取方法对绵羊血液中布氏杆菌DNA的提取效果及对PCR检测的影响。将不同浓度的疫苗株布氏杆菌加入绵羊全血中,采用3种DNA提取试剂盒和酚/氯仿法以及碘化钠法等5种方法提取模拟的绵羊血液样品中的DNA,评价所获得DNA的浓度、纯度和完整性,并采用布氏杆菌特异性PCR进行检测。同时,对各提取方法所需时间及经济成本进行了比较。结果表明,各方法均能提取获得绵羊全血中布氏杆菌DNA,3种试剂盒和碘化钠法获取布氏杆菌DNA的效果相同,而酚/氯仿法获取布氏杆菌DNA的效率最低或存在PCR抑制剂而不适合用于绵羊血液中布氏杆菌的PCR检测。碘化钠法具有耗时较短、成本低、方便的优点,是从绵羊血液中提取布氏杆菌DNA的良好方法。本研究结果为临床绵羊血液中布氏杆菌DNA提取方法的选择提供了参考。The objective of this study was to evaluate the performance of five DNA extraction methods for recovery of Brucella DNA from sheep blood and their effect on PCR detection.Different concentrations of the vaccine strain Brucella were added to the sheep blood,and the total DNA from simulated sheep blood samples was extracted using three commercial DNA extraction kits,a phenol/chloroform assay and a sodium iodide method.The concentration,purity and integrity of the obtained DNA were evaluated,and the Burcella-specific PCR was also used to detect the extracted Burcella DNA.The time and costs of each method was also evaluated.The results showed that all the methods could extract Brucella DNA from the whole blood of sheep.The three commercial kits and the sodium iodide method showed same good performance in obtaining Brucella DNA,while the conventional phenol/chloroform method was proven to be unsuitable for Brucella DNA extraction from sheep blood.The sodium iodide method had the advantages of being convenient and low-cost,and therefore was one of the best methods for Brucella DNA extraction from sheep blood.This study provided reference for selection of Brucella DNA extraction methods from clinical sheep blood.

关 键 词：绵羊 血液 布氏杆菌 DNA提取 

分 类 号：S858.26[农业科学—临床兽医学]