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A polyclonal antiserum against a recombinant viral protein combines specificity with versatility
A M Vaira1, M Vecchiati, V Masenga
1Istituto di Fitovirologia Applicata, CNR, Torino, Italy.
Journal of Virological Methods
|February 1, 1996
Summary
Researchers developed a specific antibody for tomato spotted wilt virus (TSWV) nucleoprotein. This versatile antibody works across multiple serological techniques for accurate TSWV detection.
Area of Science:
- Plant Virology
- Immunology
- Molecular Biology
Background:
- Tomato spotted wilt virus (TSWV) is a significant plant pathogen.
- Accurate and specific detection methods are crucial for disease management.
- Nucleoprotein is a key viral component for serological assays.
Purpose of the Study:
- To generate a polyclonal rabbit antiserum against the TSWV nucleoprotein.
- To evaluate the specificity and versatility of the generated antibodies.
- To assess the utility of the antibodies in various serological techniques.
Main Methods:
- Recombinant expression of TSWV nucleoprotein in E. coli.
- Production of polyclonal rabbit antiserum.
- Testing via indirect plate trapping ELISA, Western blotting, DAS-ELISA, and immunogold electron microscopy.
Main Results:
- The antiserum demonstrated high detectability and specificity against TSWV nucleoprotein.
- Antibodies showed similar titers against native nucleocapsids and recombinant fusion protein.
- Cross-reactivity was observed with groundnut ringspot virus, but not impatiens necrotic spot or groundnut bud necrosis viruses, correlating with nucleoprotein sequence similarity.
Conclusions:
- The developed antibodies are specific to TSWV nucleoprotein and versatile for multiple serological assays.
- The use of recombinant protein for immunization avoided undesired reactions from host material.
- This antibody offers a reliable tool for TSWV diagnosis and research.