Flow microsphere immunoassay-based method of virus quantitation.
A sensitive assay for adenovirus quantitation in vitro was developed using the flow microsphere immunoassay (FMIA) approach. Polystyrene microspheres were covalently coated with purified anti-adenoviral antibodies and incubated with virus-containing samples. After incubation, the samples were stained with DNA-specific fluorescent dyes, and microsphere-associated fluorescence was quantitated with a flow cytometer. The adsorption of virus to microspheres was examined under different experimental conditions. The flow cytometric assay was determined to be as accurate in detecting adenovirus as titering on 293 cells. The proposed method can be used to quantify virus in viral stocks and in biological samples.
Samoylova
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TI
College of Veterinary Medicine, Auburn University, AL, USA.
Smith
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BF
D000256:Adenoviridae / Q000276:immunology / Q000302:isolation & purification*; D000327:Adsorption; D000911:Antibodies, Monoclonal / Q000276:immunology; D001665:Binding Sites; D004926:Escherichia coli; D005456:Fluorescent Dyes / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D008863:Microspheres; D013844:Thiazoles / Q000737:chemistry; D013997:Time Factors
D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; C086329:TOTO-1 thiazole orange dye; D013844:Thiazoles