基于TaqMan探针的发菜实时荧光PCR检测方法及其应用  

Establishment and application of TaqMan probe real-time PCR method for identification of Nostoc flagelliforme

作　　者：张吉红 徐瑛 金怡伶 王佳莹 张慧丽 段维军 Zhang Jihong;Xu Ying;Jin Yiling;Wang Jiaying;Zhangg Huili;Duan Weijun(Ningbo Customs District Technical Centre,Ningbo 315012,China;Ningbo Academy of Inspection and Quarantine;Zhejiang Wanli University)

机构地区：[1]宁波海关技术中心,浙江宁波315012 [2]宁波检验检疫科学技术研究院 [3]浙江万里学院

出　　处：《植物检疫》2024年第6期38-42,共5页Plant Quarantine

基　　金：海关总署科研项目(2024HK072)。

摘　　要：本研究根据发菜组氨酸激酶基因(HK)序列设计特异性引物与荧光探针,建立了发菜实时荧光PCR检测方法。利用实时荧光PCR检测不同来源的发菜以及近似种,测定特异性及灵敏度。特异性检测结果表明,11株不同来源的发菜均为阳性扩增,与沼泽念珠藻、林氏念珠藻等其他16株念珠藻样本无交叉反应,表明该方法特异性好。灵敏度检测结果表明,本方法检测灵敏度可以达到20μL反应体系中发菜检测DNA限量为5.0 fg/μL。该方法可灵敏、快速、特异地检测鉴定发菜成分,适用于发菜及其制品的真伪鉴定。In this study,specific primer and probe were designed and selected based on partial gene sequence encoding the Nostoc flagelliforme histidine kinase(HK).A real-time PCR method was developed for identification of Nostoc flagelliforme accordingly.The N.flagelliforme samples from different sources and closely related samples were deteted by real-time PCR.The specificity and sensitivity was determined.Specificity analysis results showed that typical amplification curves were obtained for 11 N.flagelliforme samples,but not for 16 other samples such as N.paludosum,N.linckia.It indicated that the method has good specificity.The detection sensitivity was calculated to be 5.0 fg/μL in 20μL reaction system.The established real-time PCR method in this study enables rapid,sensitive and specific identification of N.flagelliforme.It is suitable for authenticity verification of N.flagelliforme and their products.

关 键 词：实时荧光PCR 发菜 检测 

分 类 号：S41[农业科学—植物保护]