Detection of double-stranded RNA by ELISA and dot immunobinding assay using an antiserum to synthetic polynucleotides
Identifieur interne : 001012 ( Istex/Curation ); précédent : 001011; suivant : 001013Detection of double-stranded RNA by ELISA and dot immunobinding assay using an antiserum to synthetic polynucleotides
Auteurs : José Aramburu [Espagne] ; Jesús Navas-Castillo [Espagne] ; Pedro Moreno [Espagne] ; Mariano Cambra [Espagne]Source :
- Journal of Virological Methods [ 0166-0934 ] ; 1991.
Abstract
An antiserum against polyinosinic-polycytidylic acid (In-Cn) was used to detect double-stranded RNA (dsRNA) by several serological techniques. DsRNA was readily detected by indirect ELISA (ELISA-I) and dot immunobinding assay (DIA). Addition of the antigen to poly-l-lysine-precoated plates and blocking with uncreamed milk powder allowed detection levels of 100 pg · ml−1 In-Cn by ELISA-I. Concentrations as low as 1 ng · ml−1 were detected by DIA using polyvinyliden difluoride (PVDF) membranes. Detection capacity with nitrocellulose membranes was 1000 times lower than with PVDF. ELISA-I and DIA enabled detection of dsRNA in enriched fractions from cucumber mosaic virus (CMV)- and citrus tristeza virus (CTV)-infected plants and from virus-infected Penicillium chrysogenum mycelium. These techniques showed similar or higher sensitivity for detection of dsRNA than separation by polyacrylamide gel electrophoresis and silver staining.
Url:
DOI: 10.1016/0166-0934(91)90002-H
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DsRNA was readily detected by indirect ELISA (ELISA-I) and dot immunobinding assay (DIA). Addition of the antigen to poly-l-lysine-precoated plates and blocking with uncreamed milk powder allowed detection levels of 100 pg · ml−1 In-Cn by ELISA-I. Concentrations as low as 1 ng · ml−1 were detected by DIA using polyvinyliden difluoride (PVDF) membranes. Detection capacity with nitrocellulose membranes was 1000 times lower than with PVDF. ELISA-I and DIA enabled detection of dsRNA in enriched fractions from cucumber mosaic virus (CMV)- and citrus tristeza virus (CTV)-infected plants and from virus-infected Penicillium chrysogenum mycelium. These techniques showed similar or higher sensitivity for detection of dsRNA than separation by polyacrylamide gel electrophoresis and silver staining.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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