Determination of infectious bursal disease virus titration and neutralization endpoints using fluorogenic staining.
An automated method for determining infectious bursal disease virus (IBDV) titration and neutralization endpoints is described. The method employs the fluorogenic ester carboxyfluorescein-diacetate (CFDA) to stain cell monolayers in 96-well plates and a fluorescence-concentration analyzer. Titration results are compared with immunofluorescence and plaque assay titers. Virus-neutralization endpoint determination is objective, and the endpoints of replicate tests were equivalent or within one dilution of variability. Tests can be automatically screened as any percentage of a positive control or any standard deviation from a negative control.
Bayyari
G R
GR
Department of Animal and Poultry Sciences, University of Arkansas, Fayetteville 72701.
Skeeles
J K
JK
Story
J D
JD
Slavik
M F
MF
D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D007243:Infectious bursal disease virus / Q000254:growth & development* / Q000276:immunology; D007668:Kidney / Q000166:cytology; D009500:Neutralization Tests; D013047:Specific Pathogen-Free Organisms; D010948:Viral Plaque Assay
D005452:Fluoresceins; C027780:carboxyfluoresceindiacetate