title: Development of reverse transcription loop-mediated isothermal amplification assay as a simple detection method of Chrysanthemum stem necrosis virus in chrysanthemum and tomato.
abstract: For a simple and rapid detection of Chrysanthemum stem necrosis virus (CSNV) from chrysanthemum and tomato, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed. A primer set designed to the genome sequences of CSNV worked most efficiently at 63°C and could detect CSNV RNA within 12min by fluorescence monitoring using an isothermal DNA amplification and fluorescence detection device. The result of a specificity test using seven other viruses and one viroid-infectable chrysanthemum or tomato showed that the assay could amplify CSNV specifically, and a sensitivity comparison showed that the RT-LAMP assay was as sensitive as the reverse transcriptase polymerase chain reaction. The RT-LAMP assay using crude RNA, extracted simply, could detect CSNV. Overall, the RT-LAMP assay was found to be a simple, specific, convenient, and time-saving method for CSNV detection.
authors.affiliation: Aichi Agricultural Research Center, 1-1 Sagamine, Nagakute, Aichi 480-1193, Japan. Electronic address: riyouji_3_suzuki@pref.aichi.lg.jp.
authors.forename: Ryoji
authors.initials: R
authors.lastname: Suzuki
authors.affiliation: Aichi Agricultural Research Center, 1-1 Sagamine, Nagakute, Aichi 480-1193, Japan.
authors.forename: Shiro
authors.initials: S
authors.lastname: Fukuta
authors.affiliation: Aichi Agricultural Research Center, 1-1 Sagamine, Nagakute, Aichi 480-1193, Japan.
authors.forename: Yuho
authors.initials: Y
authors.lastname: Matsumoto
authors.affiliation: Higashi-mikawa Agricultural Institute, 11-48 Takayama, Toyohashi, Aichi 440-0833, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Hasegawa
authors.affiliation: Aichi Agricultural Research Center, 1-1 Sagamine, Nagakute, Aichi 480-1193, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Kojima
authors.affiliation: Aichi Agricultural Research Center, 1-1 Sagamine, Nagakute, Aichi 480-1193, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Hotta
authors.affiliation: Aichi Agricultural Research Center, 1-1 Sagamine, Nagakute, Aichi 480-1193, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Miyake
mesh_terms: D031202:Chrysanthemum; D017931:DNA Primers / Q000235:genetics; D005453:Fluorescence; D018551:Solanum lycopersicum; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D010942:Plant Viruses / Q000302:isolation & purification*; D012328:RNA Viruses / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D017931:DNA Primers; D012367:RNA, Viral
keywords: Chrysanthemum; Chrysanthemum stem necrosis virus (CSNV); Reverse transcription loop-mediated isothermal amplification (RT-LAMP); Simple detection method; Tomato