A detection method based on reverse transcription loop-mediated isothermal amplification for a genetically heterogeneous plantago asiatica mosaic virus.
A detection method based on reverse transcription loop-mediated isothermal amplification for a genetically heterogeneous plantago asiatica mosaic virus.
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一种基于逆转录环介导等温扩增的遗传异质车前花叶病毒检测方法。
DOI：
10.1007/s10327-015-0599-6
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发表时间：
2015
影响因子：
1.2
通讯作者：
Namba S.
中科院分区：
文献类型：
--
作者：
Komatsu K.;Maejima K.;Fujita N.;Netsu O.;Tomomitsu T.;Arie T.;Teraoka T.;Namba S.
建立了检测车前草花叶病毒（Plantago asiatica mosaic virus，PlAMV）的逆转录环介导等温扩增（RT-LAMP）方法。根据PlAMV Li 1分离物外壳蛋白基因的中心核心区设计了一组6对引物，在65 °C下检测该分离物最有效。RT-LAMP检测特异性地检测到几个具有高水平遗传和生物学变异的PlAMV分离物，但不能检测到马铃薯X病毒（同一马铃薯X病毒属中的另一种病毒）。RT-LAMP的灵敏度是常规RT-PCR的10倍。此外，使用牙签的简单方法，使用RT-LAMP检测直接从受感染的百合叶片中检测到PlAMV，而无需提取RNA。该方法操作简便，灵敏度高，可用于PlAMV的快速检测。
A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed to detect plantago asiatica mosaic virus (PlAMV), one of the most damaging lily-infecting viruses and a member of the genusPotexvirusin the familyAlphaflexiviridae. A set of six primers was designed based on the central core region of the coat protein gene of the Li1 isolate of PlAMV, which detected the isolate most efficiently at 65 °C. The RT-LAMP assay specifically detected several PlAMV isolates with a high level of genetic and biological variation, but not potato virus X (another virus species in the samePotexvirusgenus). The sensitivity of the RT-LAMP was tenfold higher than that of conventional RT-PCR. Moreover, with a simple method using a toothpick, PlAMV was directly detected from infected lily leaves using the RT-LAMP assay without RNA extraction. This simple and highly sensitive method can be used for rapid surveys for PlAMV.