Development of Serological and Molecular Diagnostic Tools for Rose Rosette Virus

Rose rosette disease, caused by Rose rosette virus (genus Emaravirus) is a major threat to the rose industry in the U.S. The only strategy currently available for disease management is early identification and eradication of the infected plants, thereby limiting its potential spread. This requires highly sensitive serological or molecular based diagnostic tools. With the aim of developing a serological diagnostic tool, rabbit polyclonal and several mouse monoclonal antibodies have been developed, specific to the nucleocapsid protein of RRV; and has been standardized for its application in several ELISA formats including double- and triple-antibody sandwich ELISA, as well as for membrane based assays. For molecular assays, reliable and sensitive diagnostic tools including real-time RT-PCR, Loop mediated isothermal amplification (LAMP), Helicase-dependent amplification utilizing self-quenched primers (HDA-SqP) and isothermal reverse transcription-recombinase polymerase amplification (RT-exoRPA) assay has been developed and validated. The sensivity, specificity, speed and potential of the serological and molecular tools developed for lab and field based detection of Rose rosette virus will be discussed.