Advanced

TCID50 determination by an immuno dot blot assay as exemplified in a study of storage conditions of infectious pancreatic necrosis virus

Svensson, Linda LU ; Hjalmarsson, Anna and Everitt, Einar LU (1999) In Journal of Virological Methods 80(1). p.17-24
Abstract
Some cell lines are difficult to grow in confluent monolayers and, therefore, the plaque assay cannot be applied since plaques may be hard to distinguish from other blank areas of the cell monolayers. To avoid this problem a rapid and sensitive immuno dot blot TCID50 method was developed using antibodies against virus antigens to detect infection and virus production. An alternative statistical method was developed to treat the scoring data and thereby obtained a coefficient of variation of 10%. To speed up the total procedure and to increase the proliferation rate of cells grown in 96-well cell culture plates, the plates were pretreated for 4 h at 4 degrees C with growth medium obtained from cell culturing flasks containing confluent cell... (More)
Some cell lines are difficult to grow in confluent monolayers and, therefore, the plaque assay cannot be applied since plaques may be hard to distinguish from other blank areas of the cell monolayers. To avoid this problem a rapid and sensitive immuno dot blot TCID50 method was developed using antibodies against virus antigens to detect infection and virus production. An alternative statistical method was developed to treat the scoring data and thereby obtained a coefficient of variation of 10%. To speed up the total procedure and to increase the proliferation rate of cells grown in 96-well cell culture plates, the plates were pretreated for 4 h at 4 degrees C with growth medium obtained from cell culturing flasks containing confluent cell monolayers. This immuno dot blot TCID50 method was applied for a study of the infectivity maintenance upon storage of infectious pancreatic necrosis virus (IPNV). Storage of IPNV at -70 degrees C with a cryoprotective agent (10% glycerol) preserved the TCID50 level even after as many as ten cycles of freezings and thawings, whereas the infectivity decreased by four orders of magnitude after storage at 4-8 degrees C for 2 months in the salt buffer used commonly. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Viral/immunology Antigens, Animals Antibodies, Viral/*immunology Cell Division Cell Line Immunoblotting/*methods Infectious pancreatic necrosis virus/*immunology/pathogenicity Oncorhynchus mykiss Rabbits Reproducibility of Results
in
Journal of Virological Methods
volume
80
issue
1
pages
17 - 24
publisher
Elsevier
external identifiers
  • scopus:0032587829
ISSN
1879-0984
DOI
10.1016/S0166-0934(99)00018-X
language
English
LU publication?
yes
id
10971416-8fe9-483a-b198-23b9a809e420 (old id 1731058)
date added to LUP
2010-12-01 16:05:41
date last changed
2017-03-27 09:26:36
@article{10971416-8fe9-483a-b198-23b9a809e420,
  abstract     = {Some cell lines are difficult to grow in confluent monolayers and, therefore, the plaque assay cannot be applied since plaques may be hard to distinguish from other blank areas of the cell monolayers. To avoid this problem a rapid and sensitive immuno dot blot TCID50 method was developed using antibodies against virus antigens to detect infection and virus production. An alternative statistical method was developed to treat the scoring data and thereby obtained a coefficient of variation of 10%. To speed up the total procedure and to increase the proliferation rate of cells grown in 96-well cell culture plates, the plates were pretreated for 4 h at 4 degrees C with growth medium obtained from cell culturing flasks containing confluent cell monolayers. This immuno dot blot TCID50 method was applied for a study of the infectivity maintenance upon storage of infectious pancreatic necrosis virus (IPNV). Storage of IPNV at -70 degrees C with a cryoprotective agent (10% glycerol) preserved the TCID50 level even after as many as ten cycles of freezings and thawings, whereas the infectivity decreased by four orders of magnitude after storage at 4-8 degrees C for 2 months in the salt buffer used commonly.},
  author       = {Svensson, Linda and Hjalmarsson, Anna and Everitt, Einar},
  issn         = {1879-0984},
  keyword      = {Viral/immunology Antigens,Animals Antibodies,Viral/*immunology Cell Division Cell Line Immunoblotting/*methods Infectious pancreatic necrosis virus/*immunology/pathogenicity Oncorhynchus mykiss Rabbits Reproducibility of Results},
  language     = {eng},
  number       = {1},
  pages        = {17--24},
  publisher    = {Elsevier},
  series       = {Journal of Virological Methods},
  title        = {TCID50 determination by an immuno dot blot assay as exemplified in a study of storage conditions of infectious pancreatic necrosis virus},
  url          = {http://dx.doi.org/10.1016/S0166-0934(99)00018-X},
  volume       = {80},
  year         = {1999},
}