Development of polyclonal and monoclonal antibodies to Rose rosette virus nucleoprotein

Garden roses, which form the cornerstone of the multi-billion dollar landscape industry, annually generate wholesale US domestic production valued at ca. $ 400 million. Over the past few decades Rose rosette disease, caused by Rose rosette virus (RRV; genus Emaravirus), has become a major threat to the rose industry in the U.S. RRV is transmitted by wind-blown eriophyid mites, and can kill a rose within 2-3 years of infection. The long-term goal of a recently-funded USDA-NIFA-SCRI Project (which includes 17 scientists in six states) is to develop roses resistant to this virus or its mite vector. The only strategy currently available for disease management is early identification and eradication of the infected plants, thereby limiting its potential spread. Key to this effort is the development of efficient diagnostic tools to enable sensitive, rapid, user-friendly and accurate detection of the virus; currently limited to nucleic acid-based methods, such as PCR. With the aim of developing a serological diagnostic tool, rabbit polyclonal and several mouse monoclonal antibodies have been developed to bacterially-expressed nucleocapsid protein (NP) of RRV. These antibodies were evaluated against and demonstrated to detect purified bacterially-expressed NP in various serological assay formats, including antigen-coated plate and triple-antibody sandwich ELISAs, and Western blots.