Objective To establish a rapid and reliable detection method on Odontoglossum ringspot virus (ORSV), a major pathogen that seriously affects the ornamental value of orchids.
Method Primers were designed for the reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay from the conserved region in the coat protein (CP) gene of ORSV available in GenBank, and its reaction conditions optimized.
Result Using the selected primer, RT-LAMP specifically identified ORSV but not Cymbidium mosaic virus (CyMV), Bean yellow mosaic virus (BYMV), Freesia mosaic virus (FreMV) or Cucumber mosaic virus (CMV). The sensitivity of the assay was 10 times higher than that of RT-PCR, while identical in positive rate on test of 20 specimens. Furthermore, the amplification products of the newly developed method could be visually inspected using SYBR Green I without gel electrophoresis.
Conclusion The RT-LAMP assay was considered a specific, sensitive, and rapid method for ORSV detection.
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