title: Differential detection of Wheat yellow mosaic virus, Japanese soil-borne wheat mosaic virus and Chinese wheat mosaic virus by reverse transcription loop-mediated isothermal amplification reaction.
abstract: A differential detection method for three wheat viruses: Wheat yellow mosaic virus (WYMV), Japanese soil-borne mosaic virus (JSBWMV) and Chinese wheat mosaic virus (CWMV) using reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction was developed. All three primer sets, which were designed from the genome sequences of WYMV, JSBWMV and CWMV respectively, worked most efficiently at 65 °C and could detect each virus RNA within 10 min by fluorescence monitoring using an isothermal DNA amplification and fluorescence detection device. Furthermore, these primer sets showed unique annealing curves. The peak denaturing temperatures of WYMV, JSBWMV and CWMV primer sets were 87.6 °C, 84.8 °C and 86.4 °C, respectively and were clearly distinguished by the isothermal DNA amplification and fluorescence detection device. The RT-LAMP assay including all three primer sets was found to be 100 times more sensitive than RT-PCR for WYMV and JSBWMV and as sensitive as RT-PCR for CWMV. The RT-LAMP method was validated for the simultaneous detection of these viruses in wheat and barley leaves.
authors.affiliation: Agro-Environmental Division, Aichi Agricultural Research Center, 1-1 Sagamine, Yazako, Nagakute, Aichi 480-1193, Japan. shirou_fukuta@pref.aichi.lg.jp
authors.forename: Shiro
authors.initials: S
authors.lastname: Fukuta
authors.forename: Masaru
authors.initials: M
authors.lastname: Tamura
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Maejima
authors.forename: Reiko
authors.initials: R
authors.lastname: Takahashi
authors.forename: Sachiko
authors.initials: S
authors.lastname: Kuwayama
authors.forename: Takako
authors.initials: T
authors.lastname: Tsuji
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Yoshida
authors.forename: Koji
authors.initials: K
authors.lastname: Itoh
authors.forename: Hajime
authors.initials: H
authors.lastname: Hashizume
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Nakajima
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Uehara
authors.forename: Yukio
authors.initials: Y
authors.lastname: Shirako
mesh_terms: D017931:DNA Primers / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D010942:Plant Viruses / Q000235:genetics / Q000302:isolation & purification*; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D014908:Triticum / Q000821:virology*
chemical_list: D017931:DNA Primers