Kim Su Min, Choi Sun Hee
Plant Virus Genbank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University, 621 Hwarangro, Nowon, Seoul, 139-774, Korea.
Virus Genes. 2015 Dec;51(3):417-22. doi: 10.1007/s11262-015-1258-x. Epub 2015 Nov 5.
A system for simultaneous detection of two orchid-infecting viruses was developed and applied to several orchid species. The detection system involved multiplex reverse transcription-polymerase chain reaction (RT-PCR) and could simultaneously identify Cymbidium mosaic virus (CymMV) and Odontoglossum ringspot virus (ORSV) from the orchid species studied. Multiplex RT-PCR was conducted using two virus-specific primer pairs and an internal control pair of primers to amplify the CymMV and ORSV coat protein regions, and orchid 18S rDNA, respectively. For optimization of multiplex RT-PCR conditions, serial dilutions of total RNA and cDNA were performed and the detection limit of the system was evaluated. The optimized multiplex detection system for CymMV and ORSV was applied to various orchid species, including several cultivars of Doritaenopsis, Cymbidium, Dendrobium, and Phalaenopsis to test the efficacy of this method. Our results indicate that the multiplex RT-PCR detection system will be a rapid, simple, and precise diagnosis tool in a range of orchid species.
开发了一种同时检测两种感染兰花病毒的系统，并将其应用于几种兰花品种。该检测系统涉及多重逆转录-聚合酶链反应（RT-PCR），能够从所研究的兰花品种中同时鉴定出建兰花叶病毒（CymMV）和齿舌兰环斑病毒（ORSV）。使用两对病毒特异性引物和一对内部对照引物进行多重RT-PCR，分别扩增CymMV和ORSV外壳蛋白区域以及兰花18S rDNA。为优化多重RT-PCR条件，对总RNA和cDNA进行了系列稀释，并评估了该系统的检测限。将优化后的CymMV和ORSV多重检测系统应用于各种兰花品种，包括多花蝴蝶兰、大花蕙兰、石斛兰和蝴蝶兰的几个栽培品种，以测试该方法的有效性。我们的结果表明，多重RT-PCR检测系统将成为一系列兰花品种快速、简单且精确的诊断工具。
