菲律宾蛤仔及其制品的Taqman荧光PCR鉴定方法

Taqman fluorescent PCR method for rapid identification of Ruditapes philippinarumus and its products

目的建立菲律宾蛤仔成分Taqman荧光PCR定性检测方法。方法根据菲律宾蛤仔线粒体细胞色素氧化酶CoxⅠ基因中的保守基因序列设计一对菲律宾蛤仔特异性引物,通过Taqman荧光PCR法进行PCR扩增反应,对菲律宾蛤仔成分进行特异性检测。结果该方法的引物对于菲律宾蛤仔成分的特异性良好;菲律宾蛤仔DNA检出限为0.04 ng/μL,可达菲律宾蛤仔肉粉质量分数的0.001%。通过对市售菲律宾蛤仔制品的检测,该方法可检测出制品中的菲律宾蛤仔成分。结论该检测方法特异性强,灵敏度高,能够用于食品中菲律宾蛤仔成分的真实性鉴别。

Objective To establish a method of qualitative detection of Ruditapes philippinarumus by Taqman fluorescence PCR. Methods A pair of Ruditapes philippinarumus specific primers were designed according to conservative gene sequence in the CoxI gene of the mitochondrial cytochrome oxidase of Ruditapes philippinarumus. The PCR amplification reaction was carried out by Taqman fluorescence PCR method, and the specificity of the Ruditapes philippinarumus was tested. Results The specificity was good between primers and Ruditapes philippinarumus DNA. Ruditapes philippinarumus DNA detection limit was 0.04 ng/μL, which could reach 0.001%of the mass fraction of Ruditapes philippinarumus meat powder. Conclusion This method is highly specific and sensitive, which can be used for the identification of the authenticity of Ruditapes philippinarumus in the food.