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Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
Published on: May 26, 2013
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Nanoparticle-based flow virometry for the analysis of individual virions
Anush Arakelyan1, Wendy Fitzgerald, Leonid Margolis
1Program in Physical Biology, Eunice Kennedy-Shriver National Institute of Child Health and Human Development, NIH, Bethesda, Maryland 20892, USA.
The Journal of Clinical Investigation
|August 9, 2013
Summary
Flow virometry analyzes individual virus particles for antigen detection, revealing heterogeneity in human immunodeficiency virus (HIV) antigen distribution not seen in bulk analysis.
Area of Science:
- Virology
- Immunology
- Nanotechnology
Background:
- Traditional methods analyze viral antigens in bulk, masking individual particle variations.
- Flow cytometry is established for cellular analysis but not yet for individual virions.
Purpose of the Study:
- Introduce and validate "flow virometry" for single-virion antigen detection.
- Investigate the heterogeneity of human immunodeficiency virus (HIV) surface antigens.
Main Methods:
- Magnetic nanoparticles bind to virions.
- Monoclonal antibodies stain virion-bound antigens.
- Magnetic columns separate complexes.
- Flow cytometry characterizes individual virions.
Main Results:
- Flow virometry demonstrates significant antigenic heterogeneity among individual HIV-1 virions.
- Bulk analysis fails to detect this virion-level heterogeneity.
- Antigen distribution varies between HIV-1 produced by different cells.
- Different HIV-1 genotypes in the same cells show increased antigenic similarity.
Conclusions:
- Flow virometry enables single-virion antigenic analysis of viruses in bodily fluids without culturing.
- This nanotechnology is applicable to diverse viruses beyond HIV.
- Reveals previously undetectable heterogeneity in viral surface antigen composition.
