Flow cytometric detection of viruses.
Representatives from several different virus families (Baculoviridae, Herpesviridae, Myoviridae, Phycodnaviridae, Picornaviridae, Podoviridae, Retroviridae, and Siphoviridae) were stained using a variety of highly fluorescent nucleic acid specific dyes (SYBR Green I, SYBR Green II, OliGreen, PicoGreen) and examined using a standard flow cytometer equipped with a standard 15 mW argon-ion laser. The highest green fluorescence intensities were obtained using SYBR Green I. DNA viruses with genome sizes between 48.5 and 300 kb could easily be detected. The fluorescence signals of the small genome-sized RNA viruses (7.4-14.5 kb) were found at the limit of detection. No significant linear relationship could be found between genome size and the green fluorescence intensity of the SYBR Green I stained virus preparations. To our knowledge, this is the first report of detecting and discriminating between a wide range of different viruses directly using flow cytometry. This rapid and precise assay represents a new and promising tool in the field of virology.
Brussaard
C P
CP
Department of Microbiology, University of Bergen, Norway. corina.brussaard@mailroom.com
Marie
D
D
Bratbak
G
G
D052160:Benzothiazoles; D003959:Diamines; D005434:Flow Cytometry / Q000379:methods; D005456:Fluorescent Dyes; D009696:Nucleic Acids / Q000737:chemistry; D009930:Organic Chemicals; D011804:Quinolines; D011933:Reagent Kits, Diagnostic; D014780:Viruses / Q000737:chemistry*
D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009696:Nucleic Acids; D009930:Organic Chemicals; D011804:Quinolines; D011933:Reagent Kits, Diagnostic; C098022:SYBR Green I