Detection of Double-Stranded-Rna by Elisa and Dot Immunobinding Assay using an Antiserum to Synthetic Polynucleotides
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 block...
| Autores principales: | , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/4339 |
| _version_ | 1855491735509008384 |
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| author | Aramburu, Jose Navas-Castillo, J. Moreno, Pedro Cambra, Mariano |
| author_browse | Aramburu, Jose Cambra, Mariano Moreno, Pedro Navas-Castillo, J. |
| author_facet | Aramburu, Jose Navas-Castillo, J. Moreno, Pedro Cambra, Mariano |
| author_sort | Aramburu, Jose |
| collection | ReDivia |
| description | 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. |
| format | Artículo |
| id | ReDivia4339 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia43392025-04-25T14:42:53Z Detection of Double-Stranded-Rna by Elisa and Dot Immunobinding Assay using an Antiserum to Synthetic Polynucleotides Aramburu, Jose Navas-Castillo, J. Moreno, Pedro Cambra, Mariano 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. 2017-06-01T10:09:53Z 2017-06-01T10:09:53Z 1991 JUN 1991 article Aramburu, J., Navascastillo, J., Moreno, P., Cambra, M. (1991). Detection of Double-Stranded-Rna by Elisa and Dot Immunobinding Assay using an Antiserum to Synthetic Polynucleotides. Journal of virological methods, 33(1-2), 1-11. 0166-0934 http://hdl.handle.net/20.500.11939/4339 10.1016/0166-0934(91)90002-H en openAccess Impreso |
| spellingShingle | Aramburu, Jose Navas-Castillo, J. Moreno, Pedro Cambra, Mariano Detection of Double-Stranded-Rna by Elisa and Dot Immunobinding Assay using an Antiserum to Synthetic Polynucleotides |
| title | Detection of Double-Stranded-Rna by Elisa and Dot Immunobinding Assay using an Antiserum to Synthetic Polynucleotides |
| title_full | Detection of Double-Stranded-Rna by Elisa and Dot Immunobinding Assay using an Antiserum to Synthetic Polynucleotides |
| title_fullStr | Detection of Double-Stranded-Rna by Elisa and Dot Immunobinding Assay using an Antiserum to Synthetic Polynucleotides |
| title_full_unstemmed | Detection of Double-Stranded-Rna by Elisa and Dot Immunobinding Assay using an Antiserum to Synthetic Polynucleotides |
| title_short | Detection of Double-Stranded-Rna by Elisa and Dot Immunobinding Assay using an Antiserum to Synthetic Polynucleotides |
| title_sort | detection of double stranded rna by elisa and dot immunobinding assay using an antiserum to synthetic polynucleotides |
| url | http://hdl.handle.net/20.500.11939/4339 |
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