ISSN:
1573-8469
Keywords:
DAS-ELISA
;
closteroviruses
;
detection
;
serology
;
recombinant protein
Source:
Springer Online Journal Archives 1860-2000
Topics:
Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
Notes:
Abstract A polyclonal antiserum (As163) specific to grapevine leafroll associated closterovirus-3 (GLRaV-3) was developed using a recombinant coat protein expressed in E. coli from a cDNA clone identified after immunoscreening of a cDNA library. Specificity of the antiserum to GLRaV-3 was shown by Western blot and immunosorbent electron microscopy. With this antiserum, an effective double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was developed for GLRaV-3 detection. To evaluate the sensitivity of the antiserum in DAS-ELISA for virus detection, different combinations of antibodies were compared. Although best results were obtained when As163 was used for coating and a monoclonal antibody (MabNY1.1) was used as an enzyme conjugate, good results were also obtained when As163 was used both for coating and as an enzyme conjugate. Using this As163–Mab system in DAS-ELISA, we confirmed the presence of GLRaV-3 in a diverse collection of leafroll infected vines.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1023/A:1008713319294
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