中国兽医科学2011，Vol.41Issue(4)：392-396,5.
鸭疫里默氏杆菌荧光定量PCR检测方法的建立
Establishment of real-time PCR for detection of Riemerella anatipesti fer
摘要
Abstract
A pair of primers and one TaqMan probe were designed according to the conserved region of the 16 S rRNA gene of R iernerella anatipestifer (RA),and a rapid fluorescent quantitative PCR(FQ-PCR) assay was successfully established for detection of RA.In result, specificity test showed that 2 RA strains produced positive reactions,while 3 other non-RA bacterial strains produced negative reactions.The equation of the assay was Ct =-3.676 lg copies + 45.17 ,and the correlation coefficient was 0.996 and PCR efficiency of crying reached up to 93.482％.There was an excellent linear correlation during the DNA concentration from 1 copy to 1 × 108 copies.The sensitivity of the assay was 1.0× 101 copies/μL,and the variation coefficient was less than 5 ％.Comparison of RA conventional PCR with FQ-PCR for detection of the samples of the heart,liver and brain from dead duckling infected with RA, the positive rates by RA conventional PCR were 72％,84％ and 92％ respectively, but 100％ positive for all samples by FQ-PCR.Thus,this FQ-PCR assay had high sensitivity,specificity and stability,which could be used for the rapid detection,epidemiological investigation and duck product quarantine of RA infection.关键词
鸭疫里默氏杆菌/荧光定量聚合酶链反应/TaqMan探针Key words
Riemerella anatipestifer/ FQ-PCR/ TaqMan probe分类
农业科技引用本文复制引用
冯金牛,陈芳艳,晏永邦,张秀,黎丁滔,区德庆,唐秀英,陈瑞爱,王林川..鸭疫里默氏杆菌荧光定量PCR检测方法的建立[J].中国兽医科学,2011,41(4):392-396,5.基金项目
广东省科技攻关项目(2006A20301001) （2006A20301001）
