实时荧光环介导等温扩增快速检测黄曲霉毒素产生菌方法的建立  

Establishment of real-time fluorescence loop-mediated isothermal amplification method for rapid detection of aflatoxin-producing fungi

作　　者：陈晨 陶泽 王可 李晴晴 安红玉 卢增慧 李拖平 李苏红[1] CHEN Chen;TAO Ze;WANG Ke;LI Qingqing;AN Hongyu;LU Zenghui;LI Tuoping;LI Suhong(College of Food Science,Shenyang Agricultural University,Shenyang 110866,China;Central Grain Reserve Xinmin Direct Warehouse Co,Xinmin 110300,China;Central Grain Reserve Shenyang Direct Warehouse Co,Shenyang 110000,China;Liaoning Inspection,Examination&Certification Centre,Shenyang 110000,China)

机构地区：[1]沈阳农业大学食品学院,辽宁沈阳110866 [2]中央储备粮新民直属库有限公司,辽宁新民110300 [3]中央储备粮沈阳直属库有限公司,辽宁沈阳110000 [4]辽宁省检验检测认证中心,辽宁沈阳110000

出　　处：《食品与发酵工业》2025年第4期335-344,共10页Food and Fermentation Industries

基　　金：沈阳市科技局社会治理科技专项(公共安全项目)(22322339)。

摘　　要：黄曲霉毒素是一种具备强致肝损伤性、致癌性和致畸性的微生物毒素。根据产黄曲霉毒素调节基因aflR、柄曲霉素转甲基酶基因omt-1、杂色曲霉素A脱氢酶基因ver-1分别对实时荧光环介导等温扩增(loop-mediated isothermal amplification,LAMP)引物进行设计,对其建立并优化LAMP体系。结果显示,该文所建立的方法针对aflR、omt-1、ver-1基因设计的引物,黄曲霉菌DNA的检出限分别是1.0×10^(-2)、1.0×10^(-3)、1.0×10^(-4) ng/μL。此体系与黄曲霉及寄生曲霉反应成阳性,与其他菌体反应皆为阴性,该方法耗时短、设备要求低,借助恒温荧光读取设备或根据显色剂颜色变化可在60 min内迅速判定待检菌株是否为黄曲霉毒素产生菌,可满足各机构现场快速高效检测的需要。Aflatoxins are microbial toxins that are highly hepatic-inducing,teratogenic and carcinogenic.The LAMP primers were designed according to the aflatoxin-producing regulatory gene aflR,the gothramoxin transmethylase gene omt-1,and the variegated aspergilloin A dehydrogenase gene ver-1,respectively,and a real-time loop-mediated isothermal amplification(LAMP)system was established and optimized.Results showed that the detection limits of Aspergillus flavus DNA for primers designed for aflR,omt-1,and ver-1 genes were 1.0×10^(-2)、1.0×10^(-3) and 1.0×10^(-4)ng/μL,respectively.The system reacted positively with Aspergillus flavus and Aspergillus parasitica,but negatively with other bacteria,with the help of a thermostatic fluorescence reading device or according to the color change of the developer,it can quickly determine whether the strains to be tested are aflatoxins within 60 minutes.

关 键 词：黄曲霉毒素产生菌 实时荧光环介导等温扩增 快速检测 黄曲霉毒素调节基因aflR 柄曲霉素转甲基酶基因omt-1 杂色曲霉素A脱氢酶基因ver-1 

分 类 号：TS207.5[轻工技术与工程—食品科学] O657.3[轻工技术与工程—食品科学与工程]