实时荧光PCR法鉴定野生食用菌中白牛肝菌成分
实时荧光PCR法鉴定野生食用菌中白牛肝菌成分
目的建立野生食用菌中白牛肝菌(Boletus bainiugan Dentinger)成分的荧光PCR检测方法。方法根据白牛肝菌的内源转录间隔区(internally transcribed spacer, ITS)基因序列设计白牛肝菌物种的特异性引物和探针, 对样品中的靶标基因片段进行检测, 并进行物种特异性检测、稳定性检测、灵敏度检测和实际应用检测。结果通过对供试的24种食用菌、动、植物材料进行检测, 只有白牛肝菌出现特异性扩增, 说明方法具有物种特异性; 方法对白牛肝菌成分的检测灵敏度为1×10-4 ng/μL白牛肝菌DNA或0.1%(m/m)白牛肝菌粉。 结论该方法简单、灵敏、快速、准确, 能应用于野生食用菌和食品中白牛肝菌的成分检测。
ObjectiveTo establish a method for identification of Boletus bainiugan Dentinger ingredients in wild edible fungus by real-time fluorescence PCR. MethodsSpecific primers and probe were designed according to highly conserved sequence of internally transcribed spacer (ITS) gene of Boletus bainiugan Dentinger to test the target gene fragment in the samples, and the specificity, sensitivity and practical application test were carried out. ResultsThe specificity of the method was verified, and only Boletus bainiugan Dentinger had specific amplification among 24 kinds of tested edible fungus, animal and plant materials. The limit of detection was 1×10-4 ng/μL Boletus bainiugan Dentinger DNA or 0.1% (m/m) Boletus bainiugan Dentinger powder. ConclusionThis method is simple, sensitive, fast and accurate, which can be used for the identification of Boletus bainiugan Dentinger ingredients in wild edible fungus and foods.
标题：实时荧光PCR法鉴定野生食用菌中白牛肝菌成分
英文标题：Identification of Boletus bainiugan Dentinger in wild edible fungi by real-time fluorescence PCR
作者：
陈丽萍 昆明海关技术中心
赵迎春 云南省食品药品监督检验研究院
李芳 昆明海关技术中心
戚思杰 西南林业大学化学化工学院
范效英 昆明海关技术中心
丁元明 昆明海关技术中心
中文关键词:野生食用菌,白牛肝菌,实时荧光PCR,
英文关键词:wild edible fungus,Boletus bainiugan Dentinger,real-time fluorescence PCR,
发表日期：2020-04-08
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