Guidelines
CELL LINES
PLANT CELL LINES
PLANT VIRUSES
PLASMIDS
PHAGES
MICROORGANISMS
GENOMIC LIBRARIES
PROCEDURES
CATALOGUE
LABORATORY PROCEDURES FOR PLANT CELL VIRUSES
REFERENCE NO.: PVC/1998/2.04
TITLE: FULL AUTHENTICATION
INTRODUCTION
While the authenticity screen is set towards proving or establishing purity of an isolate and a preliminary test by IEM for its reaction (decoration) with a specific homologous or heterologous antiserum, a full authentication is performed to establish the relationship of the particular virus isolate with other isolates in the virus collection. This is to provide the most accurate description of a virus accession and to prevent multiple accessioning of similar if not identical viruses.
Full authentication combines information of host plant reaction and of serological assays or molecular typing tests. Usually, authentication tests performed comprise serotyping by IEM and ELISA with all antisera and related viruses available in the collection. As a result, the virus isolate under authentication is determined a separate virus, a strain of an existing virus or a serotype. Since pathovars of viruses are not amenable to serological differentiation, host plant reaction using specific genotypes expressing the disease phenotype are used for most appropriate description. When available and proven reliable, a molecular authentication is performed for viruses belonging to specific virus groups.
PROCEDURE
The combination of tests used is dependent on the virus in question, the availability of antisera and the applicability of a specific test. All virus isolates are tested by EM, IEM and ELISA (
PVC/1998/2.02 Appendix 5, PVC/1998/2.03 Appendix 2, PVC/1998/ 2.04 Appendix 2), however, the complexity and extend of tests performed depends on availability of antisera and related viruses. Data pertaining to authenticity of a virus isolate are included in the database completing the database entry.
LIST OF GLASSHOUSE AND LABORATORY PROCEDURES
PVC/1998/2.02 Appendix 5 Electron microscopy - Leaf dip assay PVC/1998/2.03 Appendix 2 Immuno Electron Microscopy PVC/1998/2.04 Appendix 2 Serotyping by ELISAGuidelines prepared for CABRI by DSMZ, 3 Feb. 1998
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