Consensus-degenerate hybrid oligonucleotide primers (CODEHOPs) for the detection of novel viruses in non-human primates

Methods. 2009 Sep;49(1):32-41. doi: 10.1016/j.ymeth.2009.05.011. Epub 2009 May 27.

Abstract

Consensus-degenerate hybrid oligonucleotide primers (CODEHOPs) have proven to be a powerful tool for the identification of novel genes. CODEHOPs are designed from highly-conserved regions of multiply-aligned protein sequences from members of a gene family and are used in PCR amplification to identify distantly-related genes. The CODEHOP approach has been used to identify novel pathogens by targeting amino acid motifs conserved in specific pathogen families. We initiated a program utilizing the CODEHOP approach to develop PCR-based assays targeting a variety of viral families that are pathogens in non-human primates. We have also developed and further improved a computer program and website to facilitate the design of CODEHOP PCR primers. Here, we detail the method for the development of pathogen-specific CODEHOP PCR assays using the papillomavirus family as a target. Papillomaviruses constitute a diverse virus family infecting a wide variety of mammalian species, including humans and non-human primates. We demonstrate that our pan-papillomavirus CODEHOP assay is broadly reactive with all major branches of the virus family and show its utility in identifying a novel non-human primate papillomavirus in cynomolgus macaques.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Conserved Sequence*
  • DNA Primers* / genetics
  • DNA, Viral / genetics
  • Humans
  • Phylogeny
  • Polymerase Chain Reaction
  • Primate Diseases / virology*
  • Primates / genetics*
  • Primates / virology*
  • Virus Diseases / virology*
  • Viruses / genetics*

Substances

  • DNA Primers
  • DNA, Viral