畜牧兽医学报2017，Vol.48Issue(3)：538-543,6.DOI:10.11843/j.issn.0366-6964.2017.03.018
实时荧光LAMP法检测转基因猪的标记基因hCD39
Real-time Fluorescence Loop-mediated Isothermal Amplification for Detection of hCD39 in Transgenic Pig
摘要
Abstract
Using ESE-Quant tube scanner testing platform,we established a real-time fluorescence loop-mediated isothermal amplification (RTF-LAMP) method for rapid detection of hCD39.The method employed a set of six specially designed primers for amplification of nucleic acid under isothermal conditions at 63 ℃ for 45 min.The amplification process of LAMP was monitored by the addition of SYBR Green Ⅰ dye prior to amplification.The fluorescence intensity of SYBR Green Ⅰ dye was used as the standard of the results.Results of the study showed that the sensitivity of the method was 3 fg · μL 1,and there was no cross-reactivity with the genes of related and unrelated detected,such as hCD46,LEA29Y.More importantly,this method could be completed testing of buccal swabs specimens of transgenic pigs.The purpose of our study was to develop a simple,economical,rapid method with good sensitivity and specificity.The method could be no damage to the living body samples for testing,suitable for quick and easy on-site operation.关键词
环介导等温扩增(LAMP)/特异性检测/hCD39Key words
loop-mediated isothermal amplification (LAMP)/specific detection/hCD39分类
农业科技引用本文复制引用
李津,刘巍,李俊超,金红岩,李刚..实时荧光LAMP法检测转基因猪的标记基因hCD39[J].畜牧兽医学报,2017,48(3):538-543,6.基金项目
国家转基因重大专项(2014ZX0801203B-002) （2014ZX0801203B-002）
国家自然科学基金(31472203) （31472203）
国家科技支撑计划(2013BAD12B05) （2013BAD12B05）
FAO/IAEA国际合作项目(17453/D32025) （17453/D32025）
