中国兽医科学2017，Vol.47Issue(5)：577-581,5.DOI:10.16656/j.issn.1673-4696.2017.05.007
犬细小病毒实时荧光环介导等温扩增检测方法的建立
Development of a real-time fluorescent loop-mediated isothermal amplification for the detection of canine parvovirus
摘要
Abstract
The special primers were designed based on the published sequence of the VP2 gene of canine parvovirus,and the fluorescent dye SYBR Green Ⅰ was added into the amplification reaction.By using Deaou-308C constant temperature fluorescence detector,we established a real-time fluorescent loopmediated isothermal amplification (LAMP)method for rapid detection of CPV.The results showed that this method had the specific amplification for CPV,while the results of control of viral nucleic acidamplification were negative,the minimum detectable sensitivity was 3.72 copies/μL virus nucleic acid.The repeatability test results showed that this method had good reproducibility.Suspected disease materials had been teseted by LAMP,and the LAMP results were compared with immuune colloidal goldtechnique and the coincidence rate was up to 90.0％.This method is further applied for the detection of canine parvovirus using the faeces from the giant panda,and the results displayed that 16 out of 84 fecal samples collected from panda base were parvovirus-positive,and the positive rate was 19％.This method is finished at a constant temperature of 63 ℃ for 45 min,easy to operate and has high sensitivity and specificity.The detecting results can be real-time detection and be automaticlly interpretated from the constant temperature fluorescence detector,which is very important for early diagnosis.关键词
犬细小病毒/实时荧光环介导等温扩增/大熊猫Key words
canine parvovirus/real-time fluorescent loop-mediated isothermal amplification (LAMP)/gaint panda分类
农业科技引用本文复制引用
陈珍容,陈进会,黄杰,龚永平,文继峰,郗立新,颜其贵..犬细小病毒实时荧光环介导等温扩增检测方法的建立[J].中国兽医科学,2017,47(5):577-581,5.基金项目
成都大熊猫繁育研究基金会项目(CFP研2012-12、CFP研2012-9) （CFP研2012-12、CFP研2012-9）
