Location: Subtropical Plant Pathology Research
Title: Development of ELISA and qPCR for Squash vein yellowing virus detection Authors
Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: April 15, 2010
Publication Date: June 1, 2010
Citation: Webster, C.G., Li, W., Kousik, C.S., Adkins, S.T. 2010. Development of ELISA and qPCR for Squash vein yellowing virus detection. Phytopathology. 100 (No. 6, supplement):S134 Technical Abstract: Watermelon vine decline caused by Squash vein yellowing virus (SqVYV) is a new and emerging disease that has caused severe losses to Florida watermelon growers in recent years. First identified in 2005, SqVYV is widely distributed in southwest and west-central Florida and has recently been found infecting several cucurbit weeds, often without inducing symptoms. Although late stage symptoms of the disease are basically diagnostic for the presence of SqVYV, earlier symptoms are not as obvious and may be confused with other causes. Thus, continued development of simple and reliable diagnostic tests for early monitoring of SqVYV in watermelon and cucurbit weeds remains important as accurate identification is the first step in management. After several unsuccessful attempts to produce specific antisera from virion preparations, peptides of the SqVYV coat protein (CP) were synthesized and used to immunize rabbits. The resulting polyclonal antisera were tested in ELISA and found to react with SqVYV but to none of the other cucurbit-infecting viruses common in Florida. A real-time PCR assay is being developed that targets the CP gene of SqVYV. Initial tests have shown the PCR assay to be sensitive and specific for SqVYV. These newly developed ELISA and real-time PCR methods for SqVYV detection are being compared to existing methods of detection including: conventional RT-PCR, tissue blots and indicator hosts on both greenhouse grown and field collected samples.