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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 qualitative Taqman fluorescent PCR assay for the detection of Ruditapes philippinarum. Methods A pair of primers was designed according to the conserved gene sequence of mitochondrial cytochrome oxidase (Cox Ⅰ) in Ruditapes philippinarum. The specific primers were used to detect the components of Ruditapes philippinarum by Taqman fluorescent PCR. Results The primers of this method were highly specific to Ruditapes philippinarum. The detection limit of DNA was as low as 0.04 ng / μL, which reached 0.001% of the mass fraction of Ruditapes philippinarum. This method detects the components of the Philippine Ruditapes philippinarum through testing of the commercially available clams in the Philippines. Conclusion The method is specific, sensitive and can be used to authenticate the components of Ruditapes philippinarum in food.