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Infectivity Assays of Human Rhinovirus-A and -B Serotypes

  • Wai-Ming LeeEmail author
  • Yin Chen
  • Wensheng Wang
  • Anne Mosser
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1221)

Abstract

Infectivity is a fundamental property of viral pathogens such as human rhinoviruses (HRVs). This chapter describes two methods for measuring the infectivity of HRV-A and -B serotypes: end point dilution (TCID50) assay and plaque assay. End point dilution assay is a quantal, not quantitative, assay that determines the dilution of the sample at which 50 % of the aliquots have infectious virus. It can be used for all the HRV-A and -B serotypes and related clinical isolates that grow in cell culture and induce cytopathic effect (CPE), degenerative changes in cells that are visible under a microscope. Plaque assay is a quantitative assay that determines the number of infectious units of a virus in a sample. After an infectious unit of virus infects one cell, the infected cell produces progeny viruses that then infect and kill a circle of adjacent cells. This circle of dead cells detaches from the dish and thus leaves a clear hole in a cell monolayer. Plaque assay works only for HeLa-adapted HRV-A and -B serotypes that can make visible plaques on the cell monolayer. Currently the end point dilution assay and plaque assay have not been developed for the newly discovered HRV-C.

Key words

MRC-5 HeLa cells End point dilution Plaque assay Infectivity TCID50 PFU 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Wai-Ming Lee
    • 1
    Email author
  • Yin Chen
    • 2
  • Wensheng Wang
    • 3
  • Anne Mosser
    • 4
  1. 1.Biological Mimetics Inc.FrederickUSA
  2. 2.Department of Pharmacology and Toxicology, School of PharmacyUniversity of ArizonaTucsonUSA
  3. 3.Global Biological DevelopmentBayer HealthcareBerkeleyUSA
  4. 4.Department of MedicineUniversity of WisconsinMadisonUSA

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