核农学报2017，Vol.31Issue(5)：868-875,8.DOI:10.11869/j.issn.100-8551.2017.05.0868
环介导等温扩增技术快速检测大西洋鲑的研究
Loop-mediated Isothermal Amplification Method (LAMP) for the Rapid Identification of Salmo salar
摘要
Abstract
To investigate the application of loop-mediated isothermal amplification (LAMP) method for salmon species identification,a set of species-specific LAMP primers was designed based on the CR fragment of the mitochondrial DNA of Salmo salar.The effects of different concentrations of dNTP Mix,Mg2 + and Betaine on LAMP results were analyzed by the single factor and orthogonal experiments.The optimal conditions were determined as follows:0.2 mM of primer SS-CR-F3 and primer SS-CR-B,1.6 mM of primer SS-CR-FIP and primer SS-CR-BIP,1.4 mM of dNTP Mix,6.0 mM of Mg2+,0.8 M of Betaine,8 U of Bst DNA polymerase in 25 μL reaction mixture.The LAMP reactions were carried out at 65℃,for 60 minutes.Under these optimized conditions,the LAMP method displayed high degrees of specificity,sensitivity and stability,with the detection limit of 0.01ng.μL-1.This method could detect S.salar either in single or mixed salmon specimen.This study provided an important technical means for rapid identification of S.salar,which could be utilized in routine inspection & quarantine and market supervision of salmon species.关键词
大西洋鲑/环等温扩增技术/物种鉴别/检测Key words
Salmo salar/loop-mediated isothermal amplification method/species identification/detection引用本文复制引用
冯俊丽,叶剑,孟璐,姜晓娜,戴志远..环介导等温扩增技术快速检测大西洋鲑的研究[J].核农学报,2017,31(5):868-875,8.基金项目
浙江省教育厅科研项目(Y201534380) （Y201534380）
