SpringerProtocols: Abstract: Titration of Human Coronaviruses, HCoV-229E and HCoV-OC43, by an Indirect Immunoperoxidase Assay

8. Titration of Human Coronaviruses, HCoV-229E and HCoV-OC43, by an Indirect Immunoperoxidase Assay

By: Francine Lambert¹, Hélène Jacomy, Gabriel Marceau, Pierre J. Talbot

Abstract

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Calculation of infectious viral titers represents a basic and essential experimental approach for virologists. Classical plaque assays cannot be used for viruses that do not cause significant cytopathic effects, which is the case for strains 229E and OC43 of human coronavirus (HCoV). An alternative indirect immunoperoxidase assay (IPA) is herein described for the detection and titration of these viruses. Susceptible cells are inoculated with serial logarithmic dilutions of samples in a 96-well plate. After viral growth, viral detection by IPA yields the infectious virus titer, expressed as “Tissue Culture Infectious Dose” (TCID₅₀). This represents the dilution of a virus-containing sample at which half of a series of laboratory wells contain replicating virus. This technique is a reliable method for the titration of HCoV in biological samples (cells, tissues, or fluids).

Affiliation(s): (1) Laboratory of Neuroimmunovirology, INRS-Institut Armand- Frappier, Laval, Québec, Canada

Book Title: SARS- and Other Coronaviruses: Laboratory Protocols

Series: Methods in Molecular Biology | Volume: 454 | Year: 2008 | Page Range: 1-10 | DOI: 10.1007/978-1-59745-181-9_8

Subject: Infectious Diseases

Key Words: human coronavirus - HCoV-229E - HCoV-OC43 - cell and tissue samples - titration - immunoperoxidase assay - TCID50

References

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