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Strain differentiation of barley yellow dwarf virus isolates using specific monoclonal antibodies in immunosorbent
1AFRC, Institute of Arable Crops Research, Rothamsted Experimental Station, Harpenden, Herts., U.K.
Abstract:
Three strain-specific monoclonal antibodies, MAC. 91, MAC. 92 and MAb. MAFF2, were compared with two polyclonal antisera for the diagnosis of barley yellow dwarf virus (BYDV) strains in immunosorbent electron microscopy (ISEM). Used at 1 micrograms ml-1, the three monoclonal antibodies specifically trapped the Rothamsted PAV, RPV and MAV-like type isolates respectively, unlike the polyclonal antisera, which showed considerable heterologous reaction. The efficiency of the monoclonal and polyclonal antibodies in trapping and detecting these isolates of BYDV were compared quantitatively using ISEM and enzyme-linked immunosorbent assay (ELISA) techniques. The test provides a simple and sensitive diagnostic method for BYDV, suitable for small numbers of samples and is useful in confirming doubtful ELISA results.
Insights
Three monoclonal antibodies accurately diagnosed barley yellow dwarf virus (BYDV) strains using immunosorbent electron microscopy (ISEM). These antibodies offer a sensitive method for BYDV detection, especially for confirming enzyme-linked immunosorbent assay (ELISA) results.
Area of Science:
- Plant Virology
- Immunology
- Diagnostic Assays
Background:
- Barley yellow dwarf virus (BYDV) is a significant pathogen affecting cereal crops worldwide.
- Accurate strain differentiation is crucial for effective disease management and breeding resistant varieties.
- Existing diagnostic methods may lack specificity or sensitivity for certain BYDV strains.
Purpose of the Study:
- To evaluate the diagnostic efficacy of three strain-specific monoclonal antibodies (MAC. 91, MAC. 92, MAb. MAFF2) against BYDV strains.
- To compare the performance of monoclonal antibodies with polyclonal antisera in diagnosing BYDV.
- To assess the utility of these antibodies in immunosorbent electron microscopy (ISEM) and enzyme-linked immunosorbent assay (ELISA).
Main Methods:
- Comparison of three monoclonal antibodies (MAC. 91, MAC. 92, MAb. MAFF2) with two polyclonal antisera.
- Application of the antibodies in immunosorbent electron microscopy (ISEM) for viral particle trapping.
- Quantitative assessment of antibody efficiency using ISEM and enzyme-linked immunosorbent assay (ELISA) techniques.
Main Results:
- Monoclonal antibodies MAC. 91, MAC. 92, and MAb. MAFF2 specifically trapped Rothamsted PAV, RPV, and MAV-like BYDV isolates, respectively.
- Polyclonal antisera exhibited significant heterologous reactions, indicating lower specificity.
- Monoclonal antibodies demonstrated high efficiency in both ISEM and ELISA for BYDV detection.
Conclusions:
- Strain-specific monoclonal antibodies provide a highly sensitive and specific diagnostic tool for identifying BYDV isolates.
- These antibodies are valuable for confirming ambiguous ELISA results and diagnosing BYDV in small sample volumes.
- The developed diagnostic method offers a simple and reliable approach for plant virologists and researchers.