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Title: Serological detection of Grapevine leafroll virus 2 using an antiserum developed against the recombinant coat protein

Author
item Ling, Kai-Shu
item ZHU, HAI-YING - CORNELL UNIVERSITY
item PETROVIC, NATASA - CORNELL UNIVERSITY
item GONSALVES, DENNIS - USDA-ARS

Submitted to: Journal of Phytopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/7/2006
Publication Date: 2/1/2007
Citation: Ling, K., Zhu, H., Petrovic, N., Gonsalves, D. 2007. Serological detection of Grapevine leafroll virus 2 using an antiserum developed against the recombinant coat protein. Journal of Phytopathology. 155:65-69.

Interpretive Summary: Grapevine leafroll associated virus-2 (GLRaV-2) is one of the important components in leafroll virus complex. Detection of GLRaV-2 has been hampered due to the lack of high quality antibody for ELISA. In this study, we employed a novel approach through recombinant DNA technology to generate a fusion coat protein for antibody production. The antibody so produced was good for plate-trapped antigen ELISA, Western blot and Immuno-electron microscopy to detect GLRaV2 in field collected samples. As more and more virus genome sequences are made available, the use of recombinant DNA approach to produce a virus-specific antibody would have broad implication in improvement for timely and sensitive virus detection.

Technical Abstract: Grapevine leafroll associated virus-2 (GLRaV-2) is one of the important components of the leafroll disease complex. The coat protein gene of GLRaV-2 was cloned into a protein expression vector pMAL-c2x and the recombinant protein, consisting of the maltose binding protein (MBP) and GLRaV-2 coat protein (CP), was expressed in Escherichia coli. The recombinant MBP-CP was used to raise a high quality antiserum. When used in Western blot analysis, the anti-MBP-CP antiserum produced specific reaction to the recombinant protein as well as to the viral coat protein of GLRaV-2. Under electron microscope, the anti-MBP-CP antibodies strongly decorated the GLRaV-2 virions. Using the newly developed antiserum, an indirect plate-trapped antigen ELISA method (PTA-ELISA) was developed and successfully implemented for virus detection in field-collected samples.