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马铃薯T病毒（potato virus T,PVT）是马铃薯的重要病害之一。为建立快速、准确、灵敏、规范的检测方法以促进马铃薯顺利出口，本研究根据PVT外壳蛋白（coat protein,CP）基因设计3组引物及TaqMan探针，通过引物筛选确定最佳引物组合，利用常规PCR方法及实时荧光PCR方法进行了特异性检测和灵敏度比较。结果表明，本研究建立的PVT实时荧光PCR检测方法特异性好，与参试的马铃薯A病毒、马铃薯X病毒、马铃薯Y病毒、马铃薯黄矮病毒、李痘病毒无交叉反应；灵敏度高，最低检测浓度可达66fg/μL。本研究构建的马铃薯T病毒实时荧光PCR检测鉴定体系，可为马铃薯繁殖材料的快速检测提供技术支持。
Abstract：Potato virus T(PVT) is one of the significant diseases on potatoes. To establish a rapid,accurate,sensitive and standardized detection method to facilitate the smooth export of potatoes,three sets of primers and TaqMan probes were designed based on the coat protein(CP) gene of PVT. The optimal primer combination was determined through primer screening. Specificity detection and sensitivity comparison were conducted using conventional PCR and real-time fluorescence PCR methods. The results showed that the real-time fluorescence PCR detection method for PVT established in this study had good specificity and no cross-reaction with the tested potato A virus,potato X virus,potato Y virus,potato yellow dwarf virus and plum pox virus. It also had high sensitivity,with the lowest detection concentration reaching 66 fg/μL. This study established a real-time fluorescent PCR detection and identification system for potato virus T,providing technical support for the rapid testing of potato reproductive materials.cent quantitative PCR,RT-qPCR)、 多重 PCR(Multiplex polymerase chain reaction,mPCR)、 数字 PCR(Digital polymerase chain reaction,dPCR)、 依赖核酸序列的扩增技术(Nucleic acid sequence-based amplification,NASBA)、 滚环扩增技术 (Rolling circle amplification,RCA)、环介导等温扩增技术(Loop-mediated isothermal amplification,LAMP)、重组酶聚合酶扩增技术 (Recombinant polymerase amplification,RPA)、交叉引物恒温扩增技术(Crossing priming isothermal amplification,CPA)等 8 种[2]. 其中实时荧光定量PCR 在植物病毒检测中应用较多,例如黄瓜绿斑驳花叶病毒 (cucumber green mottle mosaic virus,CGMMV)[3-5]、李痘病毒(plum pot virus,PPV)[6-7]、番茄黑环病毒(tomato black ring virus,TBRV)[8]、 番茄褪绿病毒(tomato chlorosis virus,ToCV)[9-10]、辣椒轻斑驳病毒(pepper mild mottle virus,PMMoV)[11]、香石竹环斑病毒(carnation ringspot virus,CRSV)[12]、玉米褪绿斑驳病毒 (maize chlorotic mottle virus,MCMV)[13]、 苹 果 茎 沟病 毒 (apple stem grooving virus,ASGV)[14-15]、 菜 豆 荚 斑 驳 病 毒 (bean pod mot tle virus,BPMV)[16]、小麦线条花叶病毒(wheat streak mosaic virus,WSMV)[17]、柑橘 衰 退 病 毒 (citrus tristeza virus,CTV)[18]、 柑橘叶斑 驳 病 毒 (citrus leaf blotch virus,CLBV)[19]、 柑 橘 褪 绿 矮 缩病 毒 (citrus chlorotic dwarf-associated virus,CCDaV)[20]、 啤酒花潜隐类病毒(hop latent viroid,HLVd)[21]等,说明该
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[22] GB/T36833—2018马铃薯X病毒检疫鉴定方法.
[23] GB/T36816—2018马铃薯Y病毒检疫鉴定方法.
基本信息:
DOI：10.19662/j.cnki.issn1005-2755.2025.06.010
中图分类号:S41-30
引用信息:
[1]李春雷,王溪桥,史琳,等.马铃薯T病毒实时荧光PCR检测方法的建立[J].植物检疫,2025,39(06):62-66.DOI:10.19662/j.cnki.issn1005-2755.2025.06.010.
基金信息:
兰州海关科技计划项目（LK-2024-004）; 海关总署科技计划项目（2024HK172）
2025-11-15
2025-11-15