实时荧光定量聚合酶链式反应快速鉴定三种鳕鱼

为鉴定常见的3种鳕科鳕鱼(大西洋鳕鱼、太平洋鳕鱼和黑线鳕),选用16S rDNA基因设计区分鳕科和非鳕科鱼类的引物,选用线粒体Cytb基因设计区分3种鳕鱼的物种特异性引物,建立实时荧光定量聚合酶链式反应(qPCR)体系,对其进行物种分析。结果表明,16S rDNA qPCR体系及3种鳕鱼品种特异性qPCR体系均具有良好的性能,结合标准曲线,可以对单一或混合样品中的目标鳕鱼进行定量检测。在模拟混合样品中目标鳕鱼的相对检测灵敏度可达0.01%。通过对13种市售样品的检测,本研究设计的qPCR体系可检测出原料及深加工产品中的鳕鱼成分。综上,所建立的qPCR体系具有很强的实用性,能满足日常检测的要求,并有望作为未来鳕鱼市场管理的检测方法。

Rapid Identification of Three Cod Species by Quantitative Real-Time Polymerase Chain Reaction

A quantitative real-time PCR assay was established for analyzing and identifing three cod species (Gadus morhua, G. macrocephalus, and Melanogrammus aeglefinus). 16S rDNA gene was used to design primers for distinguishing the Gadidae and non-Gadidae species, and the mitochondrial Cytb gene was selected for discriminating three cod species. The results showed that the RT-PCR assay systems with either 16S rDNA or species-specific gene performed well. Using the standard curves constructed for each gene, the content of target cod species could be quantitatively detected in single or mixed samples. And the relative detection limit reached 0.01% in simulated mixture samples. Through the detection of 13 commercial cod products, the qPCR system could detect the cod species in raw materials and deep-processed products. In conclusion, the methods developed in this study have strong practicability could meet the needs of routine testing, and have the possibility to be adopted as a routine method for cod market management in future.