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Optimization of in situ cellular ELISA performed on influenza A virus-infected monolayers for screening of antiviral agents

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Optimization of in situ cellular ELISA performed on influenza A virus-infected monolayers for screening of antiviral agents

Auteurs : Andrzej Myc [États-Unis] ; Marie J. Anderson [États-Unis] ; James R. Baker Jr [États-Unis]

Source :

Journal of Virological Methods [ 0166-0934 ] ; 1999.

RBID : ISTEX:54FE37A28FE22E2C7C05E90047EA3C8B9820295B

English descriptors

KwdEn :
- ELISA, Influenza A virus, MDCK cells, Plaque reduction assay, Susceptibility.
Teeft :
- Absorbance, Adsorption, Agarose, Agarose overlay medium, Agents chemother, Antiviral, Antiviral activity, Antiviral agents, Assay, Bctp, Berkowitz, Best resolution, Cell damage, Cell monolayers, Cell number, Cellular, Cellular elisa, Cellular elisa performance, Cellular elisa protocol, Elisa, Glutaraldehyde, Herpes simplex virus, Highest absorbance, Incubation, Incubation period, Incubation time, Infection medium, Initial cell concentration, Inoculum, Lactate dehydrogenase, Leahy, Levin, Mdck, Mdck cells, Monolayers, Plaque, Plaque reduction assay, Rauscher murine leukemia virus, Several hours, Subjective input, Susceptibility, Susceptibility testing, Triton, Viral, Viral inoculum, Virol, Virological, Virological methods, Virus, Virus adsorption time, Virus infection, Virus particles, Xatives.

Abstract

Abstract: Viral susceptibility testing has been traditionally performed by the plaque reduction assay (PRA) which is laborious, time consuming, relatively expensive, and requires subjective input by the reader. An in situ cellular enzyme-linked immunosorbent assay (ELISA) has been developed with the potential to overcome many of the limitations of PRA and has been applied to a variety of viruses. This study establishes the specific conditions necessary for susceptibility testing of influenza A virus to antiviral agents such as amount of inoculum size, duration of incubation, fixative type, and cell number; factors which are critical to the performance of the in situ cellular ELISA. In situ cellular ELISA was found to correlate strongly with the plaque assay (PA) (R2=0.997, P<0.002). Both assays were applied to test the susceptibility of influenza A virus to a new antiviral emulsion agent and yielded comparable data. The optimized in situ cellular ELISA can serve as a reliable assay for the rapid screening of large numbers of antiviral agents.

Url:

https://api.istex.fr/ark:/67375/6H6-TTSC529R-X/fulltext.pdf

DOI: 10.1016/S0166-0934(98)00150-5

Affiliations:

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Le document en format XML

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<term>Antiviral activity</term>
<term>Antiviral agents</term>
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<term>Bctp</term>
<term>Berkowitz</term>
<term>Best resolution</term>
<term>Cell damage</term>
<term>Cell monolayers</term>
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<term>Cellular</term>
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<term>Plaque</term>
<term>Plaque reduction assay</term>
<term>Rauscher murine leukemia virus</term>
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<term>Subjective input</term>
<term>Susceptibility</term>
<term>Susceptibility testing</term>
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<term>Virological methods</term>
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<front><div type="abstract" xml:lang="en">Abstract: Viral susceptibility testing has been traditionally performed by the plaque reduction assay (PRA) which is laborious, time consuming, relatively expensive, and requires subjective input by the reader. An in situ cellular enzyme-linked immunosorbent assay (ELISA) has been developed with the potential to overcome many of the limitations of PRA and has been applied to a variety of viruses. This study establishes the specific conditions necessary for susceptibility testing of influenza A virus to antiviral agents such as amount of inoculum size, duration of incubation, fixative type, and cell number; factors which are critical to the performance of the in situ cellular ELISA. In situ cellular ELISA was found to correlate strongly with the plaque assay (PA) (R2=0.997, P<0.002). Both assays were applied to test the susceptibility of influenza A virus to a new antiviral emulsion agent and yielded comparable data. The optimized in situ cellular ELISA can serve as a reliable assay for the rapid screening of large numbers of antiviral agents.</div>
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Optimization of in situ cellular ELISA performed on influenza A virus-infected monolayers for screening of antiviral agents

Optimization of in situ cellular ELISA performed on influenza A virus-infected monolayers for screening of antiviral agents

Source :

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri