EP0097633A1
Expired / Lapsed
EP
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Determination of the sensitivity of virus to antiviral agents
Assignee: SUNDQVIST VIVI ANNE
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Filing: 1983-06-21
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Granted: —
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Expires: 2003-06-21
Abstract
Abstract A method of determining the sensitivity of a virus to an antiviral agent is described, which method comprises replication of the virus to be determined by inoculation on cells, after which the cells are incubated after addition of different concentrations of the antiviral agent and without addition of antiviral agent. The incubated cells are disrupted and the amount of virus antigen formed in the incubation is determined immunologically in a way known per se by antigen-antibody interaction, preferably with ELISA-sandwich- technique.
Inventors
- 👤 Britta Wahren
- 👤 Johan Harmenberg
- 👤 Vivi-Anne Sundqvist
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 23:19
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/70; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/5008; G01N33/53; G01N33/569; G01N33/56983; G01N2333/00; G01N2333/005; G01N2333/01; G01N2333/03
Claims Analysis
Total Claims: 8
Claim 1
Independent Claim
226 words
A method of determining the sensitivity of a virus to an anti-viral agent through inoculation of a viral isolate thereof or a material containing such a viable virus mixed with tissue culture medium on batches of cells which permit virus replication; addition of varying concentrations of the antiviral agent to the cells; incubation of the inoculated cells, to which an anti- viral agent has been added, and of at least one batch of inoculated cells to which the antiviral agent has not been added, after which the virus is allowed to replicate; measurement of the viral replication on the cells by immunological determination of the amount of viral antigen formed through incubation; said determination including a virus antigen-antibody reaction; determination of the viral sensitivity to the antiviral agent by a comparison of the viral replication obtained at the incubation of the virus cells with addition of the antiviral agent with the viral replication obtained at the incubation of the cells without addition of the antiviral agent, characterized in that, before the immunological determination the cells are disrupted by ultrasonication, hypertonic solutions, mechanical or chemical disintegration and/or freeze-thawing in the culture medium or after removal of the supernatant, followed by addition of distilled water, thereby allowing measurement of the virus antigen content of the cells before or after occurrence of a cytopathic effect in the immunological determination.
Claim 2
Independent Claim
226 words
A method of determining the sensitivity of a virus to an anti-viral agent through inoculation of a viral isolate thereof or a material containing such a viable virus mixed with tissue culture medium on batches of cells which permit virus replication; addition of varying concentrations of the antiviral agent to the cells; incubation of the inoculated cells, to which an anti- viral agent has been added, and of at least one batch of inoculated cells to which the antiviral agent has not been added, after which the virus is allowed to replicate; measurement of the viral replication on the cells by immunological determination of the amount of viral antigen formed through incubation; said determination including a virus antigen-antibody reaction; determination of the viral sensitivity to the antiviral agent by a comparison of the viral replication obtained at the incubation of the virus cells with addition of the antiviral agent with the viral replication obtained at the incubation of the cells without addition of the antiviral agent, characterized in that, before the immunological determination the cells are disrupted by ultrasonication, hypertonic solutions, mechanical or chemical disintegration and/or freeze-thawing in the culture medium or after removal of the supernatant, followed by addition of distilled water, thereby allowing measurement of the virus antigen content of the cells before or after occurrence of a cytopathic effect in the immunological determination.
Claim 3
Dependent Claim
32 words
Method according to claim 1, characterized in that the cell disruption is carried out by removing the supernatant and freezing and a subsequent thawing of the cells after addition of distilled water.
Claim 4
Dependent Claim
37 words
Method according to claim 2, characterized in that the cells are frozen to a temperature of -5°C to -70°C, preferably -20°C, at which temperatures ice crystals form after which the cells are thawed before the immunological determination.
Claim 5
Dependent Claim
32 words
Method according to any one of claims 1-3, characterized in that the immunological determination is carried out with enzyme-linked immunosorbent assay (ELISA), e.g. with inhibition technique but preferably with a sandwich technique.
Claim 6
Dependent Claim
22 words
S. Method according to any one of claims 1-4, characterized in that the antiviral agents are added in a twofold dilution series.
Claim 7
Dependent Claim
27 words
Method according to any one of claims 1-5, characterized in that the incubation of cells with virus is interrupted when an optimal virus antigen content is obtained.
Claim 8
Dependent Claim
31 words
Method according to any one of claims 1-6, characterized in that, a virus belonging to the herpes virus family is used for inoculation, such as herpes simplex, cytomegalovirus or varicella-zoster virus.
References Cited (Backward Citations)
US4297104A
Cited
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