title: Polymerase chain reaction detection of small round-structured viruses from two related hospital outbreaks of gastroenteritis using inosine-containing primers.
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.
authors.affiliation: Department of Molecular Microbiology, University Medical School, Southampton General Hospital, United Kingdom.
authors.forename: S M
authors.initials: SM
authors.lastname: Green
authors.forename: P R
authors.initials: PR
authors.lastname: Lambden
authors.forename: Y
authors.initials: Y
authors.lastname: Deng
authors.forename: J A
authors.initials: JA
authors.lastname: Lowes
authors.forename: S
authors.initials: S
authors.lastname: Lineham
authors.forename: J
authors.initials: J
authors.lastname: Bushell
authors.forename: J
authors.initials: J
authors.lastname: Rogers
authors.forename: E O
authors.initials: EO
authors.lastname: Caul
authors.forename: C R
authors.initials: CR
authors.lastname: Ashley
authors.forename: I N
authors.initials: IN
authors.lastname: Clarke
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002139:Caliciviridae / Q000145:classification / Q000235:genetics* / Q000648:ultrastructure; D017250:Caliciviridae Infections / Q000453:epidemiology* / Q000821:virology*; D003428:Cross Infection / Q000453:epidemiology* / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004196:Disease Outbreaks; D004739:England / Q000453:epidemiology; D005243:Feces / Q000382:microbiology; D005260:Female; D005759:Gastroenteritis / Q000453:epidemiology* / Q000821:virology*; D006801:Humans; D007288:Inosine / Q000235:genetics; D008297:Male; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D004279:DNA, Viral; D012367:RNA, Viral; D007288:Inosine