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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。
Norwegian salmon (scientific name Atlantic salmon) and rainbow trout fish color and texture similar. There are often signs of misplacing some Norwegian salmon fillets in the market or the presence of Norwegian salmon with rainbow trout. In order to identify the authenticity of Norwegian salmon products, we designed two common primers for salmonids using COI gene as target gene, optimized the annealing temperature and the ratio of two primer pairs, and then used the stability test to establish a SYBR-Green real-time Fluorescent Quantitative PCR Melting Curve Method for Rapid Identification of Atlantic Salmon. The results showed that Tm values of salmonids and rainbow trout were (86.29 ± 0.172) and (83.50 ±), respectively, when the salmonidae DNA template was 100 ng, the ratio of the two pairs of universal primers was 1: 1 and the annealing temperature was 50 ℃ 0.201) ° C, and there was no significant difference in Tm values between the three test lots (P = 0.772), indicating that Atlantic salmon and rainbow trout can be identified by real-time fluorescence quantitative PCR melting curve method. The method can be used for the identification of Atlantic salmon, red salmon, silver salmon and salmon and their products. The method has the advantages of fastness, convenient operation, low cost, high accuracy and good repeatability.