摘要
旨在开发用于水产品物种鉴定的检测方法,以鉴别虹鳟(Oncorhynchus mykiss)假冒大西洋鲑(Salmo salar)的现象。根据大西洋鲑和虹鳟基因组序列相似性比对结果,在核基因肌红蛋白基因区设计了三对聚合酶链式反应扩增引物,分别为大西洋鲑特异性引物、虹鳟特异性引物和鲑鳟通用引物。在上游引物5'端标记生物素分子,使扩增产物被亲和素修饰的电极表面捕获锚定,在下游引物5'端标记二茂铁分子,使扩增产物带有电化学信号。基于以上方法构建了电化学生物传感体系,通过循环伏安信号检测特定扩增引物,从而能够鉴别样品是否来源于大西洋鲑或虹鳟。本研究构建的电化学生物传感检测方法具有准确、清洁的特点,能够用于成鱼、鱼苗、鱼卵、鱼肉及其加工制品的物种鉴定,并能够从混有其他物种DNA的样品如鱼泥中成功检测鱼肉DNA的物种来源。
This study aims to develop new method for species identification of aquatic products,in order to distinguish Atlantic salmon（Salmo salar）adulterated by rainbow trout（Oncorhynchus mykiss）. Based on genomic sequence alignment of Atlantic salmon and rainbowtrout,three pairs of polymerase chain reaction（PCR）primers in the genomic region of a nuclear gene myoglobin were designed,which wereAtlantic salmon-specific primers,rainbow trout-specific primers,and universal primers for both salmon and trout,respectively. The 5＇ endsof forward primers were labeled with biotin,thus the amplification products were able to be captured and anchored on the avidin-modifiedelectrode surfaces. And the 5＇ ends of reverse primers were labeled with ferrocene,which made the amplification products electrochemicallyactive and capable to be detected by cyclic voltammetry. The constructed electrochemical bio-sensing system accurately and safely identifiedwhether the samples were from Atlantic salmon or rainbow trout. Moreover,varied samples were applicable,including adult fish,fry,eggs,as well as processed fish products,such as fish paste containing multiple DNA sources. Therefore,the electrochemical bio-sensing detectionmethod established in this study has promising prospects in quality inspection system for aquatic products.
出处
《生物技术通报》
CAS
CSCD
北大核心
2017年第6期54-61,共8页
Biotechnology Bulletin
基金
国家科技支撑计划项目(2015BAD25B01)
关键词
大西洋鲑
虹鳟
物种鉴定
循环伏安
生物传感
Atlantic salmon
rainbow trout
species identification
cyclic voltammetry
bio-sensing
