title: Simultaneous detection of Cymbidium mosaic virus and Odontoglossum ringspot virus in orchids using multiplex RT-PCR.
abstract: 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.
authors.affiliation: Plant Virus Genbank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University, 621 Hwarangro, Nowon, Seoul, 139-774, Korea.
authors.forename: Su Min
authors.initials: SM
authors.lastname: Kim
authors.affiliation: Plant Virus Genbank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University, 621 Hwarangro, Nowon, Seoul, 139-774, Korea. paxs@swu.ac.kr.
authors.forename: Sun Hee
authors.initials: SH
authors.lastname: Choi
mesh_terms: D036022:Capsid Proteins / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D031667:Dendrobium / Q000821:virology; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D029595:Orchidaceae / Q000821:virology*; D010935:Plant Diseases / Q000821:virology*; D017863:Potexvirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017898:Tobamovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D014764:Viral Proteins / Q000235:genetics
chemical_list: D036022:Capsid Proteins; D017931:DNA Primers; D004275:DNA, Ribosomal; D012367:RNA, Viral; D014764:Viral Proteins
keywords: Cymbidium mosaic virus; Doritaenopsis; Odontoglossum ringspot virus; Orchid-infecting virus; Simultaneous detection