One-step reverse transcription loop mediated isothermal amplification assay for sensitive and rapid detection of Cucurbit chlorotic yellows virus.
A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of Cucurbit chlorotic yellows virus (CCYV). In this procedure, a set of four primers matching a total of six sequences in the coat protein gene region of CCYV was synthesized for the RT-LAMP assay using total RNA extracted from CCYV-infected melon leaf tissues, and the optimum reaction temperature and assay time were determined. The sensitivity assay showed that the virus was detectable in RT-LAMP reactions at dilutions of 1×10(-11), which was 10(5) times more sensitive than the RT-PCR assay. The RT-LAMP assay for CCYV and Sweet potato chlorotic stunt virus (SPCSV) exhibited high specificity for CCYV. This simple and sensitive method has potential for detection of CCYV in samples collected in the field.
College of Plant Protection, Henan Agricultural University, Zhengzhou 450002, China.
Zhenyue
Z
Wang
Qinsheng
Q
Gu
Hu
H
Sun
Honglian
H
Li
Bingjian
B
Sun
Xiangzhi
X
Liang
Yuan
Y
Yuan
Ruili
R
Liu
Yan
Y
Shi
D036022:Capsid Proteins / Q000235:genetics; D029383:Crinivirus / Q000235:genetics / Q000302:isolation & purification*; D019662:Cucurbitaceae / Q000821:virology; D017931:DNA Primers / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000821:virology; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
D036022:Capsid Proteins; D017931:DNA Primers; D012367:RNA, Viral
CCYV; Detection; RT-LAMP