J Med Virol. 1999 Jun;58(2):188-92.

L20B cells simplify culture of polioviruses from clinical samples.

Wood DJ1, Hull B.
Author information
1
National Institute for Biological Standards and Control, Hertfordshire, England.
dwood@nibsc.ac.uk
Abstract
Culture of polioviruses from clinical samples is the gold-standard method for virological
surveillance in the world-wide initiative to eradicate wild-type polioviruses. Two poliovirus-
sensitive cell lines of human origin were used originally by the laboratories of the World Health
Organisation (WHO) global poliovirus network. However, the cell lines used, Hep2 and RD,
also support cytopathic growth of a variety of non-poliovirus enteroviruses. This can make
detection of polioviruses in samples with mixtures of viruses difficult and time consuming. The
development of mouse cell lines that express the gene for the human cellular receptor for
polioviruses allows selective poliovirus culture, because very few non-poliovirus enteroviruses
grow in these murine cells. A WHO Collaborative Study was initiated to test one such cell line,
L20B, and to compare under routine conditions the sensitivity and selectivity of L20B cells
against RD and Hep2 cells. Five laboratories in countries endemic or recently endemic for wild
polioviruses participated. A total of 425 samples were tested prospectively in all three cell lines
and there was a clear and consistent trend for greater sensitivity for polioviruses in L20B cells.
Overall, 148/160 polioviruses were detected in L20B cells compared with 89/160 in RD and
98/ 160 in Hep2. In part, this finding was due to detection in L20B cells of polioviruses from
samples that also contained non-poliovirus enteroviruses in which the poliovirus was masked
in RD or Hep2 cells. However, L20B cells were also significantly more sensitive for poliovirus
than either RD or Hep2 cells in three of the five study laboratories. The L20B cells were
completely selective for polioviruses, as 0/89 wild type non-poliovirus enteroviruses produced
cytopathic effect in L20B cells. Finally, L20B cells provided a diagnosis of poliovirus infection
in the same time as RD and Hep2 cells from samples that contained poliovirus only, but
substantially more quickly for samples that contained another enterovirus. Taken together,
these data indicate that L20B cells simplify primary diagnosis of poliovirus from clinical
samples and as a result they have been introduced for routine use by laboratories of the WHO
global poliovirus network.
PMID:

10335869
Mikrobiyol Bul. 2002 Jul-Oct;36(3-4):301-8.
[Sensitivities of various cell cultures for the isolation of enteroviruses].
[Article in Turkish]
Ozkaya E1, Korukluoğlu G, Yalçinkaya T, Türkeri A, Atak T, Kubar A.
Author information
Abstract
In the present study, the sensitivities of HEp-2 (human epithelioma), RD (rhabdomyosarcoma)
and L20B (mouse cells, that have receptors for human polioviruses) cell cultures have been
evaluated and compared, for the isolation and identification of enteroviruses from the stool and
cerebrospinal fluid samples of patients with acute flask paralysis and aseptic meningitis, which
were examined between the years 1999-2000, in Refik Saydam Institute of Hygiene Center,
Virology, Tissue Culture and Enterovirus Laboratory. Of a total of 1663 samples, 131 viral
strains were isolated, and 120 of them were identified as enteroviruses, and 11 as adenoviruses.
The isolation rates of 48 Sabin-like polioviruses from HEp-2, RD and L20B cell lines were
found similar, as 83.3%, 87.5% and 91.6%, respectively. All of 47 Echovirus strains were
isolated from RD cells, all of 13 Coxsackie type B strains were isolated from HEp-2 cells, and
all of 12 non-polio enteroviruses were isolated from RD cells. All of 11 adenovirus strains that
have been grown in Hep-2 cells, were thought to be occasionally isolated due to the passage of
viruses to gastrointestinal tract, and excreted via stool, thus having no clinical significance for
these patients. As a result, it was concluded that, all of these three cell lines and especially
L20B were sensitive for polioviruses, RD cell line being more sensitive for Echovirus, and
HEp-2 cell line being more sensitive for Coxsackie type B virus strains.

J Med Virol. 2003 May;70(1):81-5.

Recombinant murine L20B cell line supports multiplication of group A coxsackieviruses.
Nadkarni SS1, Deshpande JM.
Author information
Abstract
Sensitive, reliable, and rapid methods of virus culture are essential for wild poliovirus isolation
and identification from stool specimens collected from cases of acute flaccid paralysis.
Recently, recombinant murine cell lines expressing human poliovirus receptor (CD155) on the
cell surface have become available. These cells are sensitive selectively to poliovirus because
the poliovirus receptor is not used by other enteroviruses. In early field studies non-polio
enteroviruses were not isolated from stool samples of cases of acute flaccid paralysis using
L20B cells. For the past 3 years, L20B cells were used extensively in our laboratory for virus
culture. The objective of the present study was to identify non-polio enteroviruses causing
cytopathic changes in L20B cells. Stool specimens collected from 1,153 cases of acute flaccid
paralysis and 2,670 apparently healthy children were tested for enteroviruses using RD and
L20B cell lines. A small number of viruses other than poliovirus causing cytopathic effect in
L20B cells were isolated. Such isolates detected in other polio network laboratories in India
were also included. The virus isolates were typed using partial VP1 nucleotide sequence
analysis and virus neutralization tests. Of the 111 viruses studied, 8 were non-enteroviruses.
Among the 103 non-polio enteroviruses, 73 were identified as group A coxsackieviruses. Of
the 30 isolates that could not be characterized, 1 remained unidentified even by sequence
analysis and 29 did not reach high titers in L20B as well as RD cells. In conclusion, coxsackie
A viruses multiply in L20B cells causing cytopathic effect. Coxsackievirus A8 and
coxsackievirus A10 were predominant among the eight coxsackie A virus types so far
identified.
Polioviruses and other enteroviruses isolated from faecal samples of patients with acute
flaccid paralysis in Australia, 1996–2004
Heath Kelly Kerri A Brussen Andrew Lawrence Elizabeth Elliot John Pearn Bruce Thorley
First published: 24 May 2006 https://doi.org/10.1111/j.1440-1754.2006.00875.x Cited by: 13

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Abstract
Background: Acute flaccid paralysis (AFP) is the most common clinical presentation of
acute poliovirus infection, occurring in 0.1–1% of infected cases. AFP surveillance has been
used world‐wide to monitor the control and eradication of circulating wild poliovirus. This
study aims to review the significance of all enteroviruses, including polioviruses, isolated from
patients with AFP in Australia between 1996 and 2004.
Methods: We undertook a retrospective review of all notified cases of AFP, aged 0–15
years and resident in Australia at the time of notification. We reviewed all available clinical
and virological data for these cases and all records of the Polio Expert Committee, which
determined the final classification for all cases.
Results: There were 335 notified cases that satisfied the case definition for AFP, 162 (48%)
of whom had at least one faecal sample tested. Enteroviruses isolated from the faeces of 26
(16%) of the 162 cases were Coxsackie A24, Coxsackie B5, enterovirus 71, enterovirus 75,
echovirus 9, echovirus 11 and echovirus 18. In addition, one or more polioviruses were isolated
from the faeces of seven patients. Six of seven polioviruses were characterised as Sabin‐like,
one was not characterised, but all were considered to be incidental isolates. Five of these cases
were classified as infant botulism, one case as transverse myelitis and one as a focal
mononeuropathy.
Conclusion: With the eradication of circulating wild polioviruses, other enteroviruses are
being more commonly identified as the cause of polio‐like illnesses. In the polio end game,
when there is increased testing for polioviruses, it is important to consider infant botulism as a
differential diagnosis in cases presenting with AFP.