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首页> 美国卫生研究院文献>The Plant Pathology Journal >Development of Multiplex PCR for Simultaneous Detection of Citrus Viruses and the Incidence of Citrus Viral Diseases in Late-Maturity Citrus Trees in Jeju Island
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Development of Multiplex PCR for Simultaneous Detection of Citrus Viruses and the Incidence of Citrus Viral Diseases in Late-Maturity Citrus Trees in Jeju Island

机译：同时检测柑橘病毒和济州岛晚熟柑桔中柑橘病毒病发病率的多重PCR方法的建立

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Satsuma dwarf virus (SDV) or Citrus mosaic sadwavirus (CiMV) were not consistently detected in RT-PCR assay with the primer sets based on gene of Japan isolates. SDV and CiMV isolates were distinctively divided into two groups based on phylogenetic analysis of PP2 gene cloned from 22 Korean isolates, and the Korean CiMV and SDV isolates shared 95.5–96.2% and 97.1–97.7% sequence identity with Japanese isolate, respectively. We developed PP2-1 primer set based on the PP2 gene sequence of Korean isolates to simultaneously and effectively detect SDV and CiMV. And CTLV-2013 and CTV-po primer sets were newly designed for detection of Citrus tatter leaf virus (CTLV) and Citrus tristeza virus (CTV), respectively. Using these primer sets, a new multiplex PCR assay was developed as a means to simultaneously detect 4 citrus viruses, CTV, CTLV, SDV, and CiMV. The degree of detection by the multiplex PCR were consistent with those of uniplex RT-PCR for detection of each of the viruses. Therefore, the new multiplex PCR provides an efficient method for detecting 4 citrus viruses, which will help diagnose many citrus plants at the same time. We verified that 35.2% and 72.1% of 775 trees in 155 orchards were infected with SDV or CiMV (SDV/CiMV) and CTV by the multiplex-PCR assay, respectively, and CTLV was not detected in any of the trees tested.
机译：使用基于日本分离株基因的引物对，在RT-PCR分析中未始终检测到萨摩矮人病毒（SDV）或柑桔花叶病毒（CiMV）。根据从22个韩国分离株中克隆的PP2基因的系统发育分析，将SDV和CiMV分离株分为两类，韩国CiMV和SDV分离株与日本分离株的序列同一性分别为95.5–96.2％和97.1–97.7％。我们根据韩国分离株的PP2基因序列开发了PP2-1引物组，以同时有效检测SDV和CiMV。并分别设计了CTLV-2013和CTV-po引物组，分别用于检测柑橘破烂叶病毒（CTLV）和柑橘三叶病毒（CTV）。使用这些引物组，开发了一种新的多重PCR分析方法，可同时检测4种柑橘病毒，CTV，CTLV，SDV和CiMV。多重PCR的检测程度与用于检测每种病毒的单重RT-PCR的检测程度一致。因此，新的多重PCR提供了一种检测4种柑橘类病毒的有效方法，这将有助于同时诊断许多柑橘类植物。我们通过多重PCR分析验证了155个果园中775棵树的35.2％和72.1％分别被SDV或CiMV（SDV / CiMV）和CTV感染，并且在任何测试的树中均未检测到CTLV。

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