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A tetrazolium-based colorimetric assay for titration of neutralizing antibodies against vaccinia virus
Year:
1998
Source of publication :
Viral Immunology
Authors :
Shoresh, Michal
;
.
Volume :
11
Co-Authors:
Ifrah, M., Department of Virology, Hebrew University, Hadassah Medical School, Jerusalem, Israel
Stienlauf, S., Department of Virology, Hebrew University, Hadassah Medical School, Jerusalem, Israel
Shoresh, M., Department of Virology, Hebrew University, Hadassah Medical School, Jerusalem, Israel
Katz, E., Department of Virology, Hebrew University, Hadassah Medical School, Jerusalem, Israel, Department of Virology, Hebrew University, Hadassah Medical School, P.O. Box 12272, Jerusalem 91120, Israel
Facilitators :
From page:
49
To page:
54
(
Total pages:
6
)
Abstract:
A colorimetric assay for titration of neutralizing antibodies against vaccinia virus was developed. The test is based on the ability of live cells in culture to reduce the yellow tetrazolium salt MTT (thiazolyl-blue), to its blue formazan derivative. Antisera from individuals vaccinated with vaccinia virus against smallpox were serially diluted, incubated with 100 plaque-forming units (PFU) of vaccinia virus for 1 hour at 37°C, and then transferred to a 96-well plate containing monolayers of B-SC-1 cells. After incubation for 3 to 4 days at 37°C, when more than 80% of the control infected cultures exhibited high degree of cytopathogenic effect, MTT was added. The absorbance of the formazan formed and extracted by dimethylsulfoxide was read at 492 nm by an automatic microplate spectrophotometer. A good correlation was found between the results obtained using this newly developed method and those of the plaque-reduction assay.
Note:
Related Files :
animal cell
Antibodies, Viral
colorimetry
formazan
smallpox vaccine
temperature
Thiazoles
Vaccination
Show More
Related Content
More details
DOI :
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
26153
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:20
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Scientific Publication
A tetrazolium-based colorimetric assay for titration of neutralizing antibodies against vaccinia virus
11
Ifrah, M., Department of Virology, Hebrew University, Hadassah Medical School, Jerusalem, Israel
Stienlauf, S., Department of Virology, Hebrew University, Hadassah Medical School, Jerusalem, Israel
Shoresh, M., Department of Virology, Hebrew University, Hadassah Medical School, Jerusalem, Israel
Katz, E., Department of Virology, Hebrew University, Hadassah Medical School, Jerusalem, Israel, Department of Virology, Hebrew University, Hadassah Medical School, P.O. Box 12272, Jerusalem 91120, Israel
A tetrazolium-based colorimetric assay for titration of neutralizing antibodies against vaccinia virus
A colorimetric assay for titration of neutralizing antibodies against vaccinia virus was developed. The test is based on the ability of live cells in culture to reduce the yellow tetrazolium salt MTT (thiazolyl-blue), to its blue formazan derivative. Antisera from individuals vaccinated with vaccinia virus against smallpox were serially diluted, incubated with 100 plaque-forming units (PFU) of vaccinia virus for 1 hour at 37°C, and then transferred to a 96-well plate containing monolayers of B-SC-1 cells. After incubation for 3 to 4 days at 37°C, when more than 80% of the control infected cultures exhibited high degree of cytopathogenic effect, MTT was added. The absorbance of the formazan formed and extracted by dimethylsulfoxide was read at 492 nm by an automatic microplate spectrophotometer. A good correlation was found between the results obtained using this newly developed method and those of the plaque-reduction assay.
Scientific Publication
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