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.
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References
Enders JF, Weller TH, Robbins FC (1949) Cultivation of the Lansing strain of poliomyelitis virus in cultures of various human embryonic tissues. Science 109:85–87
Robbins FC, Enders JF, Weller TH (1950) Cytopathogenic effect of poliomyelitis viruses in vitro on human embryonic tissues. Proc Soc Exp Biol Med 75:370–374
Robbins FC, Enders JF, Weller TH et al (1951) Studies on the cultivation of poliomyelitis viruses in tissue culture. V The direct isolation and serologic identification of virus strains in tissue culture from patients with nonparalytic and paralytic poliomyelitis. Am J Hyg 54:286–293
Robbins FC, Weller TH, Enders JF (1952) Studies on the cultivation of poliomyelitis viruses in tissue culture. II The propagation of the poliomyelitis viruses in roller-tube cultures of various human tissues. J Immunol 69:673–694
Freshney RI (2010) Culture of animal cells. A manual of basic techniques and specialized applications, 6th edn. Wiley, New Jersey
Dietz V, Andrus J, Olivé J-M et al (1995) Epidemiology and clinical characteristics of acute flaccid paralysis associated with non-polio enterovirus isolation: the experience in the Americas. Bull World Health Organ 73:597–603
Laxmivandana R, Yergolkar P, Gopalkrishna V et al (2013) Characterization of the non-polio enteroviruses infections associated with acute flaccid paralysis in South-Western India. PLoS One 8(4):e61650. doi:10.1371/journal.pone.0061650
Geraghty RJ, Capes-Davis A, Davis JM et al (2014) Guidelines for the use of cell lines in biomedical research. Br J Cancer 111:1021–1046
Pipkin PA, Wood DJ, Racaniello VR et al (1993) Characterisation of L cells expressing the human poliovirus receptor for the specific detection of polioviruses in vitro. J Virol Methods 41:333–340
Wood DJ, Hull B (1999) L20B cells simplify culture of polioviruses from clinical samples. J Med Virol 58:188–192
Nadkarni SS, Deshpande JM (2003) Recombinant murine L20B cell line supports multiplication of group A coxsackieviruses. J Med Virol 70:81–85
World Health Organization (2004) Polio laboratory manual WHO/IVB/04.10, 4th edn. WHO Department of Immunization, Vaccines and Biologicals, Geneva
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Thorley, B.R., Roberts, J.A. (2016). Isolation and Characterization of Poliovirus in Cell Culture Systems. In: Martín, J. (eds) Poliovirus. Methods in Molecular Biology, vol 1387. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3292-4_4
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DOI: https://doi.org/10.1007/978-1-4939-3292-4_4
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3291-7
Online ISBN: 978-1-4939-3292-4
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