title: Isolation and Characterization of Poliovirus in Cell Culture Systems.
abstract: The isolation and characterization of enteroviruses by cell culture was accepted as the "gold standard" by clinical virology laboratories. Methods for the direct detection of all enteroviruses by reverse transcription polymerase chain reaction, targeting a conserved region of the genome, have largely supplanted cell culture as the principal diagnostic procedure. However, the World Health Organization's Global Polio Eradication Initiative continues to rely upon cell culture to isolate poliovirus due to the lack of a reliable sensitive genetic test for direct typing of enteroviruses from clinical specimens. Poliovirus is able to infect a wide range of mammalian cell lines, with CD155 identified as the primary human receptor for all three seroytpes, and virus replication leads to an observable cytopathic effect. Inoculation of cell lines with extracts of clinical specimens and subsequent passaging of the cells leads to an increased virus titre. Cultured isolates of poliovirus are suitable for testing by a variety of methods and remain viable for years when stored at low temperature.This chapter describes general procedures for establishing a cell bank and routine passaging of cell lines. While the sections on specimen preparation and virus isolation focus on poliovirus, the protocols are suitable for other enteroviruses.
authors.affiliation: National Enterovirus Reference Laboratory, Victorian Infectious Diseases Reference Laboratory, Doherty Institute, 792 Elizabeth Street, Melbourne, VIC, 3000, Australia. bruce.thorley@mh.org.au.
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Thorley
authors.affiliation: National Enterovirus Reference Laboratory, Victorian Infectious Diseases Reference Laboratory, Doherty Institute, 792 Elizabeth Street, Melbourne, VIC, 3000, Australia.
authors.forename: Jason A
authors.initials: JA
authors.lastname: Roberts
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D006801:Humans; D011051:Poliomyelitis / Q000821:virology*; D017955:Poliovirus / Q000302:isolation & purification*
keywords: Cell bank; Cell culture; Continuous cell lines; Cryopreservation of mammalian cells; Cytopathic effect; Enterovirus; Extraction of clinical specimens; Immunofluorescence; Poliovirus; Virus isolation