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园艺学报 ›› 2005, Vol. 32 ›› Issue (02): 284-287.

• 研究论文 • 上一篇    下一篇

应用多重RT2PCR检测百合无症病毒和百合斑驳病毒

王继华1;王丽花1;丁元明2;瞿素萍1;熊丽1;唐开学1   

  1. (1 云南省农业科学院农业部花卉产品质检中心, 昆明650205; 2 云南出入境检验检疫局, 昆明650228)
  • 收稿日期:2004-07-14 修回日期:2004-11-01 出版日期:2005-04-25 发布日期:2005-04-25
  • 通讯作者: 唐开学

Detection of Lily symptomless virus and Lily mottle virus by Multiplex RT-PCR

Wang Jihua1;Wang Lihua1;Ding Yuanming2;Qu Suping1;Xiong Li1;Tang Kaixue1

  

  1. (1 Yunnan Agriculture Academy, Supervision and Testing Centre for Flower of Agriculture Ministry, Kunming 650205, China;2 Yunnan Entry-exit Inspection & Quarantine Bureau, Kunming 650228, China)
  • Received:2004-07-14 Revised:2004-11-01 Online:2005-04-25 Published:2005-04-25
  • Contact: Tang Kaixue

摘要: 根据病毒外壳蛋白基因序列, 设计了2对检测百合无症病毒(LSV) 、百合斑驳病毒(LMoV)的引物, 对扩增条件进行优化, 建立了同时检测LSV和LMoV的多重RT-PCR检测方法。此方法可特异地从带有LSV和LMoV的样品中扩增出2条带LSV (876 bp) 、LMoV (662 bp)。灵敏性测定结果表明, 该双重PCR可从稀释104 组织中检测出病毒, 具有与单一PCR相同的灵敏性。扩增产物测序表明, LSV扩增产物与其它分离物核苷酸同源性为87.8%~99.3%, LMoV扩增产物与其它分离物的同源性为90.1%~99.5%。

关键词: 多重PCR, 检测, 百合无症病毒, 百合斑驳病毒

Abstract: Two sets of specific primers were designed according to LSV and LM oV CP gene sequence,and the conditions for PCR were optimized. The multiplex PCR method which can simultaneously detect the two lily viruses was developed. It was showed that all samples which infected LSV and LMoV could be amplified by multiplex PCR, yielding two specific bands of LSV (876 bp) and LMoV (662 bp). The two viruses were detected from dilutions of 104 by multiplex RT-PCR and RT-PCR, respectively. Sequence analysis on the amplified products show that the nucleotide sequences homology of LSV was 87.8% -99.3% compared with
sequences of other isolates, and LMoV was 90.1%-99.5%.

Key words: Multiplex PCR, Detection, Lily symptomless virus, Lily mottle virus

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