基于线粒体16S rRNA基因的鹅肉源性成分鉴别方法研究

Identification Method for Goose Meat Origin Ingredients Based on Mitochondrial 16S rRNA Gene

研究旨在建立基于线粒体16S rRNA基因的鹅源性成分鉴别方法。试验以鹅源性DNA为阳性模板,以猪、牛、羊、鸽、鹌鹑、火鸡、鸡和鸭等8个物种DNA为干扰模板的混合模板,设计筛选出鹅特异性引物,进行PCR和荧光定量PCR(qPCR)反应,并将鹅肉DNA模板浓度按101~1088个梯度进行稀释,检测方法灵敏度。结果显示:所设计的引物仅对鹅肉DNA有特异性扩增,对鹅以外的其他8个物种均没有扩增;当鹅肉DNA模板稀释104倍,PCR扩增条带仍然清晰;当稀释倍数达到107时,仍有较好的扩增曲线,且Ct值小于35。研究表明,建立的畜禽肉中鹅源性成分PCR和qPCR鉴别方法不仅具有良好的特异性,而且具有较高的灵敏性,为食品中鹅源性成分的鉴别提供了新途径。

The study was aimed to establish an indentification method for goose meat origin ingredients based on mitochondrial 16S RNA gene,which could identify the components of goose origin.Mitochondrial DNA 16S rRNA gene of pig,cow,sheep,turkey,chickens,ducks,goose,pigeon and quail were used as target sites and the goose specific primers were designed and screened.Nine animals DNA were used as templates for conventional PCR and fluorogenic quantitative PCR(qPCR),and the goose DNA template was diluted in eight gradients,including 101 times to108 times,for detecting the sensitivity.The results showed that the designed and screened primers only amplified the DNA of goose meat,and did not amplify the DNA of other 8 animals except geese.Sensitivity detection showed that when the DNA template of goose meat was diluted 104 times,PCR amplification bands were still clear.When the dilution ratio reached 107,there was still a good amplification curve,and the Ct value was less than 35.It's suggested that the PCR and qPCR methods established in this study not only had good specificity,but also had high sensitivity,which would provide a new way for the identification of goose-derived components in food.