| 易汪雪1, 宋绍祎1, 吴东妮2, 代欢欢3, 杨翠云1, 于 翠1*.单管多重RT-PCR同时检测大豆种子中三种检疫性植物病毒[J].植物保护,2016,42(5):113-117. |
| 单管多重RT-PCR同时检测大豆种子中三种检疫性植物病毒 |
| Simultaneous detection of three quarantine plant viruses from soybean seeds by multiplex RT-PCR in single PCR tube |
| 投稿时间：2015-11-06 修订日期：2015-12-24 |
| DOI： |
| 中文关键词: 菜豆荚斑驳病毒 烟草环斑病毒 番茄环斑病毒 多重RT-PCR 同时检测 |
| 英文关键词:Bean pod mottle virus Tobacco ringspot virus Tomato ringspot virus multiplex RT-PCR simultaneous detection |
| 基金项目:上海市科技兴农重点攻关项目［沪农科攻字(2012)第2-8号］；上海出入境检验检疫局科研项目（HK005-2014） |
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| 中文摘要: |
| 我国大豆年进口量持续增长，快速、准确地检测进境大豆种子中可能携带的病原物是防止检疫性有害生物入境传播和扩散的有效手段。菜豆荚斑驳病毒(BPMV)、烟草环斑病毒(TRSV)和番茄环斑病毒(ToRSV)均是被大豆种子携带的检疫性有害生物。本研究建立了在单一PCR管中同时检测这3种检疫性植物病毒及大豆内源基因Bd-30 K的多重RT-PCR方法。研究结果表明，所建立的多重RT-PCR方法具有较好的特异性和灵敏度，检测含BPMV、TRSV和ToRSV RNA的最低浓度分别为0.45、0.0093和0.004 ng/μL，从大豆种子中同时检测3种病毒的最低RNA浓度为0.58 ng/μL。 |
| 英文摘要: |
| The quantity of soybean seeds imported were continually increased in recent years. Rapid detection of seed-borne pathogens is essential to efficiently prevent pest invading and spreading. Bean pot mosaic virus (BPMV), Tobacco ringspot virus (TRSV) and Tomato ringspot virus (ToRSV) are the quarantine pest which could be carried by the soybean seeds. We developed the multiplex RT-PCR approach for simultaneous detection of BPMV, TRSV, ToRSV and one endogenous gene Bd 30K in the single PCR tube from soybean seeds. The results indicated that good specificity and sensitivity for simultaneous detection were obtained. The limit detection of RNA with BPMV, TRSV and ToRSV were 0.45 ng/μL, 0.009 3 ng/μL and 0.004 ng/μL, respectively. The limit detection of RNA with three viruses mixture was 0.58 ng/μL |
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