Taqman探针荧光PCR检测鲨鱼源性成分

Detection of Shark Derived Materials by Taqman Probe Fluorescent PCR

摘要:

针对鲨鱼产品掺杂造假增多而缺乏有效鉴定技术的情况，作者根据GenBank中鲨鱼基因的线粒体DNA（mtDNA）序列，使用分子生物学软件Primer Express 2.0设计了一套特异性引物和探针，用来建立Taqman探针荧光PCR检测鲨鱼源性成分的方法。结果表明，对13份已鉴定为鲨鱼鱼翅的样品进行检测，全部出现特异性扩增曲线，阴性对照没有荧光增长。方法特异性强，对市场购买的12份鲨鱼源性样品及9份其他鱼类样品进行检测，12份鲨鱼样品均出现荧光扩增曲线，而非鲨鱼样品均未出现荧光增长；方法灵敏度较高，可检测到的最低质粒拷贝数量级为10拷贝/μL；方法快速准确，操作简便，重复性好，稳定可靠，可应用于市场上鱼翅等常见鲨鱼源性产品的真假鉴定。

Abstract:

As the problem of adulteration of shark products worsens and the effective detection method lacks, this study designed a set of primers and probe based on the mitochondria DNA sequences of shark published in GenBank using the software of Primer Express 2.0, so as to establish a method of Taqman probe fluorescent PCR for the detection of shark derived materials. Results showed that 13 samples that had been identified as shark fins all displayed the amplified curves. The specificity of this method was good with all of 12 shark derived samples indicating the existence of shark DNA and with no amplification curves for the other 9 samples not derived from shark species. The detection is sensitive and the minimum detectable plasmid copy number was 10 copies/μL. It is also rapid and simple and the results have good repeatability and stability. Therefore, the method can be applied to the identification of shark derived products in marketplaces, such as fins.