中国食品学报2023，Vol.23Issue(12)：196-208,13.DOI:10.16429/j.1009-7848.2023.12.021
GⅠ和GⅡ诺如病毒RAA可视化快速检测方法的建立
Establishment of a Visual Recombinase Aided Amplification(RAA)Method for Rapid Detection of GⅠ and GⅡ Noroviruses
摘要
Abstract
Norovirus(NoV)is one of the major pathogens causing acute gastroenteritis diseases worldwide and is highly susceptible to outbreak transmission,increasing medical and economic burden.Objective:To develop a novel detection method for NoV by recombinase aided amplification(RAA).Methods Based on the cDNA sequences corresponding to the GⅠ and GⅡ NoV detection targets specified in ISO TS 15216-2-2013 and GB 4789.42,the optimal RAA primers and probes were designed and screened,and their specificity for other common food-bome diarrhea viruses was determined;the shortest detection time,reaction procedure,and reaction system were determined by optimization,and the analysis of this detection system was performed.The shortest detection time,reaction procedure and reaction system were optimized,and the sensitivity of the assay system for the detection of NoV reference plasmids and real samples was analyzed,thus establishing a rapid visualization method for the detection of GⅠ and GⅡ NoV RAA.The optimized reaction procedure can shorten the detection time to about 10 min and reduce the reaction cost by two-thirds,and the sensitivity of the method can reach 10-2 ng/μL for the reference plasmid and 1 ng/μL for the real sample.The two NoV assays established are specific,sensitive,simple,rapid and visualized,and provide a good basis for future rapid NoV detection.关键词
诺如病毒(NoV)/重组酶介导等温扩增(RAA)/可视化/快速检测Key words
Norovirus(Nov)/recombinase mediated isothermal amplification(RAA)/visualization/rapid detection引用本文复制引用
林志伟,杨艳歌,吴占文,王帅,李涛,李红娜,袁飞..GⅠ和GⅡ诺如病毒RAA可视化快速检测方法的建立[J].中国食品学报,2023,23(12):196-208,13.基金项目
国家重点研发计划项目(2022YFF1100900,2022YFF0607900) （2022YFF1100900,2022YFF0607900）
