绿脓杆菌焦磷酸测序检测方法的建立  

Identification of Pseudomonas aeruginosa Using Pyrosequencing Assays

作　　者：张太翔[1] 孙军[1] 赵晗[1] 李秀勇 孙涛 刘文鹏[1] 田国宁[1] 韩亮[1] 

机构地区：[1]潍坊出入境检验检疫局,山东潍坊261041 [2]烟台出入境检验检疫局,山东烟台260010 [3]山东检验检疫局,山东青岛250023

出　　处：《中国畜牧兽医》2012年第8期53-56,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家标准化管理委员会2010年国家标准制修订项目(2010B071.2)

摘　　要：本试验旨在利用绿脓杆菌的序列信息分析和焦磷酸测序技术,建立一种快速、简单检测绿脓杆菌的方法。从培养的绿脓杆菌中提取DNA,PCR扩增目的基因片段,采用焦磷酸测序技术(pyrosequencing technology,PSQ)针对保守核苷酸区段的测序分析。通过焦磷酸测序后获得的序列信息与已知的目的基因的序列比对,能进一步确证毒株的序列信息为绿脓杆菌。利用焦磷酸测序能快速获取核酸的序列信息,可为毒株及早确证奠定基础。In this study,we reported a new method of pyrosequencing-based sequence analysis to detect Pseudomonas aeruginosa,which was a rapid simple method.After extracting DNA from cultured cells,Pseudomonas aeruginosa were preliminarily determined by PCR on a specific sequence of the exotoxin A gene which containing conserved fragment respectively.Then the results obtained by PCR were further validated via the pyrosequencing method,and the sequences were demonstrated to specific of Pseudomonas aeruginosa.By conventional sequencing,the sequences results were 100% in agreement with pyrosequencing.Comparing the sequence got by pyrosequencing with the known sequence,you would find the sequence was Pseudomonas aeruginosa.This method was accurate,fast,and could be used efficiently for identifying the Pseudomonas aeruginosa.

关 键 词：绿脓杆菌 PCR 焦磷酸测序 鉴定 

分 类 号：S816.79[农业科学—饲料科学]