Development of immunocapture reverse transcription loop-mediated isothermal amplification for the detection of tomato spotted wilt virus from chrysanthemum.
An immunocapture reverse transcription loop-mediated isothermal amplification (IC/RT-LAMP) was developed for the detection of tomato spotted wilt virus (TSWV) from chrysanthemum. This method enabled sensitive, reproducible and specific detection of TSWV from chrysanthemum plants. In the RT-LAMP method, TSWV genomic RNA could be amplified under isothermal (65 degrees C) conditions within 1 h. The resulting amplicons were detected by the measurement or observation of the turbidity of the reaction mixture without gel electrophoresis. IC/RT-LAMP was 100 times more sensitive than IC/RT-PCR.
Fukuta
Shiro
S
Aichi Prefecture Agricultural Research Center, 1-1 Sagamine, Yazako, Nagakute, Aichi 480-1193, Japan. shirou_fukuta@pref.aichi.lg.jp
Ohishi
Kazushi
K
Yoshida
Keiko
K
Mizukami
Yuko
Y
Ishida
Akira
A
Kanbe
Michio
M
D001483:Base Sequence; D031202:Chrysanthemum / Q000821:virology*; D008969:Molecular Sequence Data; D009391:Nephelometry and Turbidimetry; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017865:Tospovirus / Q000235:genetics / Q000302:isolation & purification*
D012367:RNA, Viral