鳕鱼成分的实时荧光PCR检测方法

Detection of Gadiformes by Real-time PCR Assay

研究建立了鳕鱼成分的实时荧光PCR检测方法。依据鳕鱼的16S rRNA基因序列设计一对PCR引物及探针,对12种鳕鱼样品和71种非鳕鱼动植物样品进行实时荧光PCR检测,结果显示,只有12种鳕鱼样品产生荧光信号,其他非鳕鱼样品均不产生荧光信号,实验表明此方法具有特异性,其检测限为0.01 ng/μl鳕鱼DNA和0.01%鳕鱼肉粉。对市售的鳕鱼样品进行实际样品检测,均能很好地检出鳕鱼成分。此法特异性强,灵敏度高,可作为鳕鱼成分鉴定的检测方法。

A rapid and sensitive TaqMan real-time PCR assay were developed for the detection of gadiformes. For high gadiformes specificity, the primers and the probe were designed on the basis of the mitochondrial 16S rRNA gene. The detection limit of this real-time PCR assay was 0. 01 ng/μl DNA concentration and 0.01% （W/W） gadiformes meat powder. To confirm the specificity of these methods, genomic DNA samples from 71other species were examined; no amplification products were detected. Thus, the method shows high sensitivity and may be applicable as specific tools for the detection of trace amounts of gadiformes in commercial food products.