摘要
建立一种可同时快速检测红薯、木薯、马铃薯、玉米源性成分的多重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)方法。分别以红薯g3pdh基因、木薯g3pdh基因、马铃薯UGPase基因、玉米zSSIIb基因为靶基因设计特异性引物和TaqMan探针,以18S rRNA基因为内参基因,建立多重real-time PCR方法,开展方法学验证,并对不同掺入比例模拟样品和实际淀粉样品进行检测。结果显示,该方法具有高通量、特异性强、灵敏度高等优点。与15种非目标源性均无交叉反应;对目标DNA的检测灵敏度可达到3×10^(-3) ng/μL,且具有良好的线性关系和扩增效率;对淀粉样品的检出限可达0.1%,对50份实际样品进行检测,结果与参比方法一致,说明建立的多重real-time PCR法可用于食用淀粉种类掺假鉴别检测。
A multiplex real-time polymerase chain reaction(PCR)method for simultaneous and rapid detection of components derived from sweet potato,cassava,potato,and corn was established.The g3pdhs gene of sweet potato and cassava,the UGPase gene of potato and the zSSIIb gene of corn were used as target genes to design specific primers and TaqMan probe,and the 18S rRNA gene was used as the internal reference gene.Methodological validation was performed,and the proposed method was applied to simulated samples with different mixing ratios and actual starch samples.The results showed that this method was high-throughput,sensitive and specific.No cross-reaction was found with 15 non-target sources.The detection sensitivity for template DNA was 3×10^(-3) ng/μL,and the proposed method had a good linear relationship and high amplification efficiency.The detection limit for edible starch samples was 0.1%.The results of the PCR method for 50 actual samples were consistent with those of the reference method,indicating that this method could be used for the detection of common adulterants in edible starches.
作者
范维
高晓月
董雨馨
刘虹宇
李贺楠
赵文涛
郭文萍
FAN Wei;GAO Xiaoyue;DONG Yuxin;LIU Hongyu;LI Henan;ZHAO Wentao;GUO Wenping(China Meat Research Center,Beijing Academy of Food Sciences,Beijing 100068,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2024年第1期210-216,共7页
Food Science
基金
北京市科技计划项目(Z221100007122010)
“十三五”国家重点研发计划重点专项(2018YFC1603400)
北京市优秀人才培养资助青年骨干个人项目(2017754154700G101)。
关键词
多重实时聚合酶链式反应
食用淀粉
木薯
红薯
马铃薯
玉米
multiplex real-time polymerase chain reaction
edible starch
cassava
sweet potato
potato
corn
作者简介
第一作者:范维(1989-)(ORCID:0000-0001-6122-4727),女,高级工程师,硕士,研究方向为食品科学与工程。E-mail:fw837093515@163.com;通信作者:郭文萍(1970-)(ORCID:0000-0002-9677-0397),女,高级工程师,学士,研究方向为食品安全。E-mail:956920027@qq.com。
