猪痢疾短螺旋体荧光PCR检测方法的建立与初步应用.pdfVIP

Chin曲e 10‘unal 0{ Vet回inary Medicine 中国兽医杂志 2014年(第so4第6期 9 猪病疾短螺旋体荧光PCR检测方法的 建立与初步应用 张伟\高志强，李宁2 ，谷强\ 凌凤俊t ，乔彩霞1 ，蒲静，张利峰1 ， 汪琳，吴丹1 ，柏亚锋3 ，安健3 ，刘环\赖平安\张鹤晓 (l北京出入编检验检疫局，北京通州 101113 ; 2. 中国兽医药品监察所，北京海淀 100081 ; 3.北京农学院，北京 昌平 102206) 摘要:在对设计合成的引物和6-carboxy-l1 uo陀scein (6-FAM)荧Jr.:.J;_ 标记的Taq Man水解探针进行筛选的基础上. 泊过反 山体系优化. 建立了猪躏疾短拟旋体荣先PCIl 险ìlJ方法， 特异性、敏感性、重复性及i 临床样品的俭i则如果均农~Y1 建立的方法 快速、特异、灵敏，整个检测过程(包摇-样品的前处理时间)可在4h 内在戚。 边立的方法大大缩短 n家病原的检测周期，并为 临床诊断提供快速、敏感、特异性强的俭测方法。 关键词:猪躏疾;猪躏疾短螺旋体;荧光PCR 中图分类号: S852.65.1 文献标，悄地 文章编号:0529-6∞5(2014)06-ω9-04 Development and Application of TaqMan Real-time PCR for Brachyspira hyodysenteriae Z H ANG Wei , GAO Zhi -qiang 1 , LI Ning2 , GU Qiang , LlNG Feng-jun , QIAO Cai-xia I , P U Jing 1 , ZHANG Li-feng 1 , WANG Lin , WU Da n , BA I Y a-duo , AN J ianl , LlU Hu a n , LA I P ing-an , Z H ANG H e-xiao ( I.Beijing Entry- Exit Inspection and Quarantine ßureau. Beijing 101113 ,China; 2.China Institute of Veterinary Drug Control. Beijing 1 仪泊8 1 •China; 3. Beijing University of A伊culture ， ß eijing 102206 ,China) Abstract: Based on the scrt:ening of the designed prilllers and TaqMan hydrolysis type probes labeled with 6- carboxy-f1 uoresce- in (6-FAM) reporter dyes. thi s rl:earch r1eveloped a real-time PC R detection method for Brachyspira hyodysenteriae by means of op - timazati on of reaction conditions. The sensitivity, specifì cit y飞 repeatability IInd detection c1inical samples of th e real -time P CR were cxamin erl resp ectively. Th e result~ showed Ihat th e method were rap id , specifi c and sen sitive. The assays coul d be fìnished in 4 hou附 (including the p陀treatmenl of samples). Th ( developed l1lethod shortens the detecting period of swin