新冠病毒的现场可视化LAMP检测及其冷链食品分析应用  

LAMP Assay for On-site Detection of SARS-CoV-2 and Its Application in Cold-chain Food Analysis

作　　者：刘楠 澹台玮 王芳 徐秦峰 LIU Nan;TANTAI Wei;WANG Fang;XU Qinfeng(Shaanxi University of Science&Technology,School of Food and Biological Engineering,Xi’an 710021,China)

机构地区：[1]陕西科技大学食品与生物工程学院,陕西西安710021

出　　处：《现代食品科技》2022年第1期88-93,共6页Modern Food Science and Technology

基　　金：国家重点研发计划项目(2019YFC1606302);国家自然科学基金项目(22074085)。

摘　　要：建立了一种基于环介导等温扩增技术(Loop-mediated Isothermal Amplification,LAMP)和核酸分子"光开关"[Ru(phen)_(2)dppz]^(2+)相结合的新型冠状病毒高灵敏可视化检测方法。分别采用保温杯代替实验室恒温装置,采用蓝光手电代替蓝光透射仪,实现了新冠病毒的现场快速可视化LAMP检测。针对新型冠状病毒N基因设计特异性序列引物,采用[Ru(phen)_(2)dppz]^(2+)分子光开关的特性检测LAMP扩增产物,结果可以借助蓝光手电直接通过肉眼观察是否出现红色荧光进行判定。该方法可检测到单个拷贝数的新型冠状病毒基因片段,并具有高度特异性,检验快速,整个过程仅需40min即可目视观察结果。对新冠病毒假病毒人工污染的模拟冷链食品进行检测,结果与目前新型冠状病毒检测金标准实时荧光定量PCR法吻合度为100%。该方法仅需保温杯和蓝光手电,可以作为实时荧光PCR方法的补充,为食品新冠病毒的现场筛查提供快速高效的技术支持。A highly sensitive visualization method for SARS-CoV-2 detection,based on loop-mediated isothermal amplification(LAMP)and molecular light switch[Ru(phen)_(2)dppz]^(2+),was established.In this design,insulation cups replaced laboratory thermostats and blue flashlights replaced blue transilluminators,to realize the rapid on-site visual LAMP detection of SARS-Co V-2.Specific primers were designed for the N gene of SARS-CoV-2.LAMP amplification products were detected by[Ru(phen)_(2)dppz]^(2+)with molecular light switch characteristics.Red fluorescence could be directly detected by naked eye using blue light flashlight.Single copy number SARS-CoV-2 gene fragments were detected with high specificity.The detection was rapid,requiring only 40 minutes to visually observe the results.The LAMP detection results for food samples artificially contaminated with SARS-CoV-2 pseudovirus were 100%consistent with the current gold-standard real-time fluorescent quantitative PCR method for SARS-CoV-2 detection.This method requires only insulation cups and blue flashlight,and can be used to supplement the real-time fluorescent PCR method to provide a fast and efficient on-site screening of food for SARS-CoV-2.

关 键 词：环介导等温扩增技术 新型冠状病毒 [Ru(phen) dppz] 现场可视化LAMP检测 冷链食品 

分 类 号：TS207.3[轻工技术与工程—食品科学]