Abstract
During the past years increasing incidences of influenza A zoonosis have made it of uppermost importance to possess methods for rapid and precise identification and characterisation of influenza A viruses. We present here a convenient one-step RT-PCR method that will amplify full-length haemagglutinin (HA) and neuraminidase (NA) directly from clinical samples and from all known subtypes of influenza A. We applied the method on samples collected in September 2003 from a Danish flock of mallards with general health problems and by this a previously undescribed influenza A subtype combination, H5N7, was identified. The HA gene showed great sequence similarity to the highly pathogenic avian influenza A virus (HPAIV) A/Chicken/Italy/312/97 (H5N2); however, the cleavage site sequence between HA1 and HA2 had a motif typical for low pathogenic avian influenza viruses (LPAIV). The full-length NA sequence was most closely related to the HPAIV A/Chicken/Netherlands/01/03 (H7N7) that infected chickens and humans in the Netherlands in 2003. Ten persons with direct or indirect contact with the Danish mallard ducks showed signs of influenza-like illness 2-3 days following the killing of the ducks, but no evidence of influence infections was detected. To our knowledge this is the first report of an H5N7 influenza A virus.
MeSH terms
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Animals
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DNA, Complementary
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DNA, Viral / chemistry
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DNA, Viral / metabolism
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Denmark
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Ducks / virology
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Genes, Viral
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Hemagglutinin Glycoproteins, Influenza Virus / genetics
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Humans
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Influenza A virus / classification*
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Influenza A virus / isolation & purification*
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Influenza in Birds / virology*
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Influenza, Human / virology*
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Molecular Sequence Data
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Neuraminidase / genetics
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Phylogeny
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RNA, Viral / analysis
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RNA, Viral / isolation & purification
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Reverse Transcriptase Polymerase Chain Reaction
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Sequence Analysis, DNA
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Sequence Homology
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Viral Proteins / genetics
Substances
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DNA, Complementary
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DNA, Viral
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Hemagglutinin Glycoproteins, Influenza Virus
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RNA, Viral
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Viral Proteins
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Neuraminidase
Associated data
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GENBANK/AY531006
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GENBANK/AY531007
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GENBANK/AY531008
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GENBANK/AY531009
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GENBANK/AY531010
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