title: [Development of a real-time RT-PCR for detection of equine influenza virus].
abstract: Equine influenza is a highly contagious respiratory disease in horses caused by influenza A viruses. In this work a real-time RT-PCR for fast and sensitive diagnosis of equine influenza viruses (EIV) targeting a highly conserved region of the matrix gene was developed. In addition two RT-PCR methods for the amplification of large parts of the matrix- and HA gene were adapted for molecular-epidemiological characterization of viruses. The primers of the real-time RT-PCR had homologies of 99.4% to EIV- and 97.7% to all influenza A viral sequences, whereas the minor groove binder (MGB) probe showed homologies of 99.3% and 99.6%, respectively. These high values allow application of the assay for influenza viruses in other species. Using 20 equine, 11 porcine and 2 avian samples, diagnostic suitability of the assay was confirmed. High specificity for influenza viruses was shown both experimentally and by software simulation. The assay analytical sensitivity was at 10(2)-10(3) copies of RNA and 10(0)-10(1) copies of DNA, respectively. This allows virus detection also in circumstances of minor viral shedding. All amplified EIV sequences were classified phylogenetically within the known lineages.
authors.forename: S
authors.initials: S
authors.lastname: Aeschbacher
authors.forename: E
authors.initials: E
authors.lastname: Santschi
authors.forename: V
authors.initials: V
authors.lastname: Gerber
authors.forename: H P
authors.initials: HP
authors.lastname: Stalder
authors.forename: R G
authors.initials: RG
authors.lastname: Zanoni
mesh_terms: D000818:Animals; D001717:Birds; D004285:Dogs; D006734:Horse Diseases / Q000175:diagnosis* / Q000821:virology; D006736:Horses; D009980:Influenza A virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D061985:Madin Darby Canine Kidney Cells; D009976:Orthomyxoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D014763:Viral Matrix Proteins / Q000235:genetics; D017201:Virus Shedding / Q000235:genetics
chemical_list: D014763:Viral Matrix Proteins