title: Complete genome amplification of equine influenza virus subtype 2.
abstract: This work reports a method for rapid amplification of the complete genome of equine influenza virus subtype 2 (H3N8). A ThermoScript reverse transcriptase instead of the avian myeloblastosis virus reverse transcriptase or Moloney murine leukemia virus reverse transcriptase was used. This enzyme has demonstrated higher thermal stability and is described as suitable to make long cDNA with a complex secondary structure. The product obtained by this method can be cloned, used in later sequencing reactions or nested-PCR with the purpose of achieving a rapid diagnosis and characterization of the equine influenza virus type A. This detection assay might be a valuable tool for diagnosis and screening of field samples as well as for conducting molecular studies.
authors.affiliation: Catedra de Virología, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Buenos Aires, Argentina. sguazza@fcv.unlp.edu.ar
authors.forename: G H
authors.initials: GH
authors.lastname: Sguazza
authors.forename: N A
authors.initials: NA
authors.lastname: Fuentealba
authors.forename: M A
authors.initials: MA
authors.lastname: Tizzano
authors.forename: C M
authors.initials: CM
authors.lastname: Galosi
authors.forename: M R
authors.initials: MR
authors.lastname: Pecoraro
mesh_terms: D000818:Animals; D016384:Consensus Sequence; D017124:Conserved Sequence; D016679:Genome, Viral; D053123:Influenza A Virus, H3N8 Subtype / Q000145:classification / Q000235:genetics*; D009976:Orthomyxoviridae Infections / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D012194:RNA-Directed DNA Polymerase; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid
chemical_list: D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase