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Subcellular fractions in rubella immunoassays
European Journal of Clinical Microbiology
|February 1, 1984
Summary
This study fractionated rubella virus-infected cells, finding viral antigens in various cellular components. Enriched fractions, particularly cellular membranes, proved useful for rubella virus antibody detection, enhancing IgM test specificity.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Rubella virus infection requires sensitive diagnostic methods.
- Understanding viral antigen localization within host cells is crucial for developing improved immunoassays.
Purpose of the Study:
- To investigate the distribution of rubella virus antigens within infected cells.
- To evaluate the utility of cellular fractions as antigen sources for rubella virus-specific antibody detection.
Main Methods:
- Fractionation of rubella virus-infected cells using differential and sucrose gradient centrifugation.
- Analysis of antigen distribution across various cellular fractions.
- Solid-phase immunoassay development using isolated cellular membranes and pellets for detecting rubella virus-specific IgG and IgM antibodies.
Main Results:
- Rubella virus antigens were detected in all cellular fractions, with enrichment in the 100,000 x g pellet and enriched fractions of rough endoplasmic reticulum and smooth cellular membranes.
- These enriched fractions demonstrated utility in solid-phase immunoassays for rubella virus antibodies.
- While comparable to semi-purified preparations for IgG detection, the fractions were inferior for IgM assays, though non-specific binding was identified and addressed.
Conclusions:
- Cellular membrane fractions from rubella virus-infected cells are valuable sources for rubella virus antigen detection.
- Specific adjustments, such as using fractions from both infected and non-infected cells, can improve the specificity of IgM immunoassays for rubella virus.