热带作物学报2025，Vol.46Issue(7)：1745-1753,9.DOI:10.3969/j.issn.1000-2561.2025.07.021
齿兰环斑病毒RT-RPA荧光实时检测体系的构建与应用
Construction and Application of RT-RPA Fluorescence Real-time Detection System for Odontoglossum Ringspot Virus
摘要
Abstract
Odontoglossum ringspot virus(ORSV),one of the major pathogens affecting orchids,seriously degrade the ornamental and economic value of orchids by causing leaf chlorosis,mottling,and floral deformities.To enable early-stage diagnosis,this study developed a rapid reverse transcription-recombinase polymerase amplification(RT-RPA)assay targeting ORSV.Primers and a fluorescent probe were designed based on conserved regions of the ORSV coat protein(CP)gene.Specificity,sensitivity and field applicability of the assay were systematically validated in this study.Results identified F1/R1 as the optimal primer pair,enabling amplification within 20 minutes at 41 ℃.The method exhibited high specificity,distinguishing ORSV from Cymbidium mosaic virus and Cucumber mosaic virus,and achieved a de-tection limit of 2.8×10-5ng/μL total RNA.Practical verification result of 20 Phalaenopsis field samples showed 100％concordance with TaqMan qPCR results.The RT-RPA assay would provide a rapid,sensitive,and field-deployable tool for ORSV surveillance,supporting timely disease management and sustainable orchid cultivation.Its simplicity and reliability make it particularly valuable for large-scale screening in industrial orchid production systems.关键词
兰花/齿兰环斑病毒/反转录-重组酶聚合酶扩增(RT-RPA)/检测Key words
Orchidaceae/Odontoglossum ringspot virus(ORSV)/reverse transcription-recombinase polymerase ampli-fication(RT-RPA)/detection分类
农业科技引用本文复制引用
吴洁秋,庄华才,谭志勇,李早文,范俊强,傅海平,徐匆..齿兰环斑病毒RT-RPA荧光实时检测体系的构建与应用[J].热带作物学报,2025,46(7):1745-1753,9.基金项目
2022年度广东省基础与应用基础研究基金(区域联合基金)项目(No.2022A1515140182). （区域联合基金）
