One-step reverse transcription loop-mediated isothermal amplification for the rapid detection of cucumber green mottle mosaic virus.
Cucumber green mottle mosaic virus (CGMMV) has caused serious damage to Cucurbitaceae crops worldwide. The virus is considered one of the most serious Cucurbitaceae quarantine causes in many countries. In this study, a highly efficient and practical one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed for the detection of CGMMV. The total RNA or crude RNA extracted from watermelon plants or seeds could be detected easily by this RT-LAMP assay. The RT-LAMP assay was conducted in isothermal (63°C) conditions within 1h. The amplified products of CGMMV could be detected as ladder-like bands using agarose gel electrophoresis or visualized in-tube under UV light with the addition of a fluorescent dye. The RT-LAMP amplification was specific to CGMMV, as no cross-reaction was observed with other viruses. The RT-LAMP assay was 100-fold more sensitive than that of reverse-transcription polymerase chain reaction (RT-PCR). This is the first report of the application of the RT-LAMP assay to detect CGMMV. The sensitive, specific and rapid RT-LAMP assay developed in this study can be applied widely in laboratories, the field and quarantine surveillance of CGMMV.
Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Department of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, China.
Jin-yu
JY
Li
Qi-wei
QW
Wei
Yong
Y
Liu
Xin-qiu
XQ
Tan
Wen-na
WN
Zhang
Jian-yan
JY
Wu
Miriam Karwitha
MK
Charimbu
Bai-shi
BS
Hu
Zhao-bang
ZB
Cheng
Cui
C
Yu
Xiao-rong
XR
Tao
D019662:Cucurbitaceae / Q000821:virology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D017898:Tobamovirus / Q000235:genetics / Q000302:isolation & purification*; D014773:Virology / Q000379:methods*
Cucumber green mottle mosaic virus; RT-LAMP; Seed; Watermelon