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Polymerase chain reaction detection of small round-structured viruses from two related hospital outbreaks of gastroenteritis using inosine-containing primers

Polymerase chain reaction detection of small round-structured viruses from two related hospital outbreaks of gastroenteritis using inosine-containing primers
Polymerase chain reaction detection of small round-structured viruses from two related hospital outbreaks of gastroenteritis using inosine-containing primers
Two outbreaks of gastroenteritis in the UK which occurred nine days apart at Lymington and Southampton hospitals were investigated. The clinical and epidemiological features of both outbreaks were characteristic of small round-structured virus (SRSV) infection with rapid onset of diarrhoea and/or nausea and vomiting and propagation of the outbreaks by secondary spread. SRSV particles were observed by immune electron microscopy (EM) in 60% of faecal samples from both outbreaks and no other pathogens were detected. The index case for the second outbreak was a patient who was admitted with diarrhoea and vomiting after being discharged from Lymington hospital during the first outbreak. The possibility that the two outbreaks were caused by the same strain of SRSV was investigated by the polymerase chain reaction (PCR). New inosine-containing PCR primers were designed to amplify the RNA polymerase region of SRSV cDNA from genetic groups I and II. The PCR using the group II primers achieved a higher detection rate for SRSVs in faecal samples (68% of samples positive from both outbreaks) than immune EM. SRSVs were not detected using the group I primers or using conventional degenerate PCR primers. The nucleotide sequences of PCR amplicons from both outbreaks were identical providing molecular epidemiological evidence for the involvement of a single SRSV strain. Comparison of the RNA polymerase region of this virus with the equivalent regions of genetic group I (69.4-75.0% amino acid identify) and genetic group II (88.9-100% amino acid and 77.1-88.1% nucleotide identity) SRSVs revealed that the causative SRSV was a distinct member of genetic group II.
calicivirus, epidemiology, nucleotide sequence, electron microscopy
0146-6615
197-202
Green, Steve M.
c11018d2-e940-4424-a0e3-7d844e5d286e
Lambden, Paul R.
4fcd536e-2d9a-4366-97c6-386e6b005698
Deng, Yu
d0f1af1d-6428-4b11-8fcb-ffef9f8c40f8
Clarke, Ian N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b
Lowes, J. Andrew
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Lineham, Sarah
5cd6ee51-c490-4154-baca-16a3b6ab9c81
Bushell, Julie
e573a9d1-0e6a-4196-887c-e8a8b7e0d61a
Rogers, John
bb38733b-d2ff-4027-94e0-fe54099c04d9
Caul, E. Owen
177391c9-26e2-4c94-a459-d9353c2daa61
Ashley, Charles R.
f5151218-e9e0-4bc5-9dfd-bac2b04a0e31
Green, Steve M.
c11018d2-e940-4424-a0e3-7d844e5d286e
Lambden, Paul R.
4fcd536e-2d9a-4366-97c6-386e6b005698
Deng, Yu
d0f1af1d-6428-4b11-8fcb-ffef9f8c40f8
Clarke, Ian N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b
Lowes, J. Andrew
9bc30920-0f7b-49ab-a5f2-6d9dedb967b6
Lineham, Sarah
5cd6ee51-c490-4154-baca-16a3b6ab9c81
Bushell, Julie
e573a9d1-0e6a-4196-887c-e8a8b7e0d61a
Rogers, John
bb38733b-d2ff-4027-94e0-fe54099c04d9
Caul, E. Owen
177391c9-26e2-4c94-a459-d9353c2daa61
Ashley, Charles R.
f5151218-e9e0-4bc5-9dfd-bac2b04a0e31

Green, Steve M., Lambden, Paul R., Deng, Yu, Clarke, Ian N., Lowes, J. Andrew, Lineham, Sarah, Bushell, Julie, Rogers, John, Caul, E. Owen and Ashley, Charles R. (1995) Polymerase chain reaction detection of small round-structured viruses from two related hospital outbreaks of gastroenteritis using inosine-containing primers. Journal of Medical Virology, 45 (2), 197-202. (doi:10.1002/jmv.1890450215). (PMID:7775939)

Record type: Article

Abstract

Two outbreaks of gastroenteritis in the UK which occurred nine days apart at Lymington and Southampton hospitals were investigated. The clinical and epidemiological features of both outbreaks were characteristic of small round-structured virus (SRSV) infection with rapid onset of diarrhoea and/or nausea and vomiting and propagation of the outbreaks by secondary spread. SRSV particles were observed by immune electron microscopy (EM) in 60% of faecal samples from both outbreaks and no other pathogens were detected. The index case for the second outbreak was a patient who was admitted with diarrhoea and vomiting after being discharged from Lymington hospital during the first outbreak. The possibility that the two outbreaks were caused by the same strain of SRSV was investigated by the polymerase chain reaction (PCR). New inosine-containing PCR primers were designed to amplify the RNA polymerase region of SRSV cDNA from genetic groups I and II. The PCR using the group II primers achieved a higher detection rate for SRSVs in faecal samples (68% of samples positive from both outbreaks) than immune EM. SRSVs were not detected using the group I primers or using conventional degenerate PCR primers. The nucleotide sequences of PCR amplicons from both outbreaks were identical providing molecular epidemiological evidence for the involvement of a single SRSV strain. Comparison of the RNA polymerase region of this virus with the equivalent regions of genetic group I (69.4-75.0% amino acid identify) and genetic group II (88.9-100% amino acid and 77.1-88.1% nucleotide identity) SRSVs revealed that the causative SRSV was a distinct member of genetic group II.

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More information

Published date: February 1995
Keywords: calicivirus, epidemiology, nucleotide sequence, electron microscopy
Organisations: Faculty of Medicine

Identifiers

Local EPrints ID: 352655
URI: https://eprints.soton.ac.uk/id/eprint/352655
ISSN: 0146-6615
PURE UUID: 6938d585-0129-49d0-9703-bd743d71dab0
ORCID for Ian N. Clarke: ORCID iD orcid.org/0000-0002-4938-1620

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Date deposited: 17 May 2013 09:34
Last modified: 06 Jun 2018 13:19

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Contributors

Author: Steve M. Green
Author: Paul R. Lambden
Author: Yu Deng
Author: Ian N. Clarke ORCID iD
Author: J. Andrew Lowes
Author: Sarah Lineham
Author: Julie Bushell
Author: John Rogers
Author: E. Owen Caul
Author: Charles R. Ashley

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