摘要
建立TaqMan实时荧光PCR快速检测粮食霉变主要霉菌的方法。根据引发粮食霉变的霉菌属5.8S rDNA的ITS序列来设计通用引物和探针,一次实时荧光PCR扩增同时检测黄曲霉和寄生曲霉,与SN/T 2582-2010中普通PCR方法进行比较,验证方法的特异性和灵敏度;通过建立粮食霉变模型并进行霉菌检测,进一步验证方法的准确性。结果表明,所设计的实时荧光PCR引物探针具有良好的特异性和灵敏性,灵敏度达到10-2 ng/μl。所建立的实时荧光PCR快速检测粮食主要霉菌的方法,快速简便准确,适用于引发粮食霉变的黄曲霉和寄生曲霉的检测。
A TaqMan real-time PCR method for rapid and high sensitivity detection of the main fungus in grain was established.The general primers and probes were designed according to the ITS sequence of 5.8S rDNA of main fungus causing grain mildew,and the real-time PCR amplification was used to detect both Aspergillus flavus and A.parasiticus.Compared with conventional PCR method in SN/T 2582-2010,the specificity and sensitivity of the method were verified,and the accuracy of the method was verified by establishing a grain mildew model and detecting the fungus.Results showed that the primers and probe of this method have good specificity and sensitivity,with a sensitivity of 10-2 ng/μl.The method is suitable for the rapid detection of Aspergillus flavus and A.parasiticus causing grain mildew.
作者
冯优妍
杨爱馥
郑秋月
万超
曹际娟
FENG You-yan;YANG Ai-fu;ZHENG Qiu-yue;WAN Chao;CAO Ji-juan(Dalian University of Technology Food College,Dalian 116034,China;Dalian Customs District,Dalian 116001,China)
出处
《粮食与饲料工业》
CAS
2020年第4期1-3,9,共4页
Cereal & Feed Industry
基金
科技部“十三五”国家重点研发计划项目(2017YFC1600803-3)
辽宁省自然科学基金项目(20180551271)。
关键词
实时荧光PCR
粮食霉变
检测
黄曲霉
寄生曲霉
特异性
灵敏度
TaqMan real-time PCR
grain mildew
detection
Aspergillus flavus
A.parasiticus
specificity
sensitivity
