Adaptation of the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay for the determination of virus-neutralizing antibodies using the virus-neutralization assay.
The tetrazolium salt MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] has been widely used for bioassays. Herein is described the use of the MTT dye with a virus-neutralization (VN) assay to titer infectious bursal disease virus (IBDV)-neutralizing antibodies. A standard VN assay using chicken embryo fibroblasts (CEFs) and IBDV was used for the assessment of IBDV-neutralizing antibodies. The percent of CEF killing due to IBDV was quantitated using MTT, and the absorbance (A) data were used to calculate the VN antibody titer. This method of calculation offers the expression of VN titer in terms of units of activity per unit of volume.
Haddad
E E
EE
EMBREX, Inc., Research Triangle Park, North Carolina 27709.
Whitfill
C E
CE
Ricks
C A
CA
Fredericksen
T
T
Rowe
D
D
Owen
L
L
Baldridge
A
A
Murray
L
L
Thoma
J A
JA
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D018175:Birnaviridae Infections / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens / Q000276:immunology* / Q000821:virology; D005347:Fibroblasts; D007243:Infectious bursal disease virus / Q000276:immunology*; D009500:Neutralization Tests / Q000662:veterinary*; D011201:Poultry Diseases / Q000276:immunology* / Q000821:virology; D012680:Sensitivity and Specificity; D013778:Tetrazolium Salts; D013844:Thiazoles
D000914:Antibodies, Viral; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue