Detection of Cymbidium mosaic virus in Protocorm-Like Bodies in Dendrobium Sonia using One-Step RT-PCR

Dendrobium orchids are clonally mass propagated by tissue culture techniques. However, Cymbidium mosaic virus is prevalent in vegetatively propagated Dendrobium spp. Therefore, a sensitive virus detection method is an important requirement for the production of CymMV-free orchid plants and control of virus spreading in the orchid industry. The presence of CymMV was screened in in vitro PLBs of Dendrobium Sonia using a one-step RT-PCR with specific primers to amplify a 858 bp fragment of a CymMV coat protein gene and flanking regions. Results were compared with those obtained from a conventional indirect ELISA. Only 22% samples showed the presence of CymMV in indirect-ELISA as compared to 78% in RT-PCR. Thus, the one-step RT-PCR appears to be a more sensitive method than indirect-ELISA for detecting CymMV in PLBs.