基于SYBR-Green熔解曲线快速鉴别挪威三文鱼及其制品的方法  

A Rapid Identification Method of the Norwegian Salmon Slices Based on SYBR Green I Real-time PCR Combined with Melting Curve Analysis

作　　者：李新光[1,2] 赵峰[1] 马丽萍[1,2] 孙永[1] 周德庆[1] 

机构地区：[1]中国水产科学研究院黄海水产研究所农业部海洋资源可持续利用重点开放实验室,山东青岛266071 [2]上海海洋大学食品学院,上海201306

出　　处：《中国食品学报》2014年第3期170-176,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：中央级公益性科研院所基本科研业务费专项资金(20603022013018)

摘　　要：挪威三文鱼(学名大西洋鲑)和虹鳟的鱼肉颜色和纹理相似。在市场上常存在将一些挪威三文鱼片错贴标签或以虹鳟冒充挪威三文鱼的现象。为了鉴别挪威三文鱼制品的真伪,以COI基因为靶基因,设计2对鲑科鱼类的通用引物,优化退火温度和两对引物的浓度比值,然后做稳定性试验,建立基于SYBR-Green实时荧光定量PCR熔解曲线快速鉴别大西洋鲑的方法。试验结果显示,当鲑科鱼类DNA模板100 ng,两对通用引物的浓度比值1∶1、退火温度50℃时,大西洋鲑和虹鳟鱼的Tm值分别为(86.29±0.172),(83.50±0.201)℃,且3个检验批次的Tm值没有显著性的差异(P=0.772),表明大西洋鲑和虹鳟可通过实时荧光定量PCR熔解曲线法进行鉴别。该方法可用于大西洋鲑与红鲑、银鲑以及大马哈鱼及其制品的鉴别,具有快速、操作方便、成本低廉、准确度高和重复性好的特点。In order to authenticate commercial frauds of Atlantic salmon（Salmo salar） products by being mislabeled or substituted with rainbow trout（Oncorhynchus mykiss）,a rapid identification method of the Norwegian salmon slices was established based on SYBR Green I Real-time PCR combined with melting curve analysis.In this experiment,COI was taken as target gene and two pairs of universal primers for salmons were designed.Then,the annealing temperatures and the ratio of two pairs of universal primers were optimized.It showed that the salmon species identification was most significantly affected by the ratio of two pairs of universal primers,followed by annealing temperatures.The commercial Atlantic salmon products can be easily differentiated from rainbow trout products by melting curve analysis when initial templates amount of salmons was added about 100ng,the two salmons＇ universal primer concentration ratio was 1∶1,annealing temperature was 50 ℃.In addition,The repeatability experiments were well performed（P=0.772）,（86.29±0.172）℃ for Atlantic salmon and（83.50 ±0.201）℃ for rainbow trout,respectively.Further studies suggested that the established method also can be used for identification of Atlantic salmon among sockeye salmon,white salmon and silver salmon.The results showed that this method was simple,low cost,accurate and sensitive with a good reproducibility.

关 键 词：大西洋鲑 虹鳟 真伪鉴别 实时荧光定量PCR 熔解曲线法 

分 类 号：S917.4[农业科学—水产科学]