Phylogenetic analysis of a highly conserved region of the polymerase gene from 11 coronaviruses and development of a consensus polymerase chain reaction assay

Charles B. Stephensen, Donald B. Casebolt, Nupur N. Gangopadhyay

Research output: Contribution to journalArticlepeer-review

102 Scopus citations

Abstract

Viruses in the genus Coronavirus are currently placed in three groups based on antigenic cross-reactivity and sequence analysis of structural protein genes. Consensus polymerase chain reaction (PCR) primers were used to obtain cDNA, then cloned and sequenced a highly conserved 922 nucleotide region in open reading frame (ORF) 1b of the polymerase (pol) gene from eight coronaviruses. These sequences were compared with published sequences for three additional coronaviruses. In this comparison, it was found that nucleotide substitution frequencies (per 100 nucleotides) varied from 46.40 to 50.13 when viruses were compared among the traditional coronavirus groups and, with one exception (the human coronavirus (HCV) 229E), varied from 2.54 to 15.89 when compared within these groups. (The substitution frequency for 229E, as compared to other members of the same group, varied from 35.37 to 35.72.) Phylogenetic analysis of these pol gene sequences resulted in groupings which correspond closely with the previously described groupings, including recent data which places the two avian coronaviruses-infectious bronchitis virus (IBV) of chickens and turkey coronavirus (TCV)-in the same group [Guy, J.S., Barnes, H.J., Smith L.G., Breslin, J., 1997. Avian Dis. 41:583-590]. A single pair of degenerate primers was identified which amplify a 251 bp region from coronaviruses of all three groups using the same reaction conditions. This consensus PCR assay for the genus Coronavirus may be useful in identifying as yet unknown coronaviruses. Copyright (C) 1999 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)181-189
Number of pages9
JournalVirus Research
Volume60
Issue number2
DOIs
StatePublished - Apr 1999
Externally publishedYes

Keywords

  • Consensus PCR
  • Coronavirus
  • Polymerase gene

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Virology

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