One-step reverse transcription loop-mediated isothermal amplification assay for rapid detection of Cymbidium mosaic virus
2011
Lee, Meng-Shiou | Yang, Meng-Ja | Hseu, You-Cheng | Lai, Guan-Hua | Zhang, Wende | Hsu, Yau-Heiu | Lin, Ming-Kuem
Cymbidium mosaic virus (CymMV) is the most prevalent orchid virus. A single-tube one-step betaine-free reverse transcription (RT) loop-mediated isothermal amplification (LAMP) assay was developed for the rapid and easy detection of orchid-infecting CymMV. Five sets of primers were designed based on the conserved regions among various virus isolates. The specificity and the sensitivity of the assay were then evaluated using the RT-LAMP reaction. Within 1h under isothermal conditions at 60°C the target viral gene was amplified successfully. This RT-LAMP assay was found to be quick, specific, sensitive and easy to perform assay that involved only one step and was simpler to carry out than alternative approaches. Thus this assay is an alternative for the rapid and easy detection of CymMV in orchids. This is first time that a RT-LAMP method for the detection of an orchid virus has been described.
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