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      Airborne transmission of influenza A/H5N1 virus between ferrets.

      Science (New York, N.Y.)

      Virus Shedding, Air Microbiology, Amino Acid Substitution, Animals, Antiviral Agents, pharmacology, Containment of Biohazards, Disease Models, Animal, Female, Ferrets, Hemagglutinin Glycoproteins, Influenza Virus, chemistry, genetics, immunology, metabolism, Humans, Immune Sera, Influenza A Virus, H5N1 Subtype, drug effects, pathogenicity, physiology, Influenza in Birds, epidemiology, virology, Influenza, Human, transmission, Molecular Sequence Data, Mutagenesis, Site-Directed, Mutation, Orthomyxoviridae Infections, Oseltamivir, Pandemics, Poultry, RNA Replicase, Reassortant Viruses, Receptors, Virus, Respiratory System, Reverse Genetics, Serial Passage, Sialic Acids, Viral Proteins, Virulence, Virus Replication

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          Abstract

          Highly pathogenic avian influenza A/H5N1 virus can cause morbidity and mortality in humans but thus far has not acquired the ability to be transmitted by aerosol or respiratory droplet ("airborne transmission") between humans. To address the concern that the virus could acquire this ability under natural conditions, we genetically modified A/H5N1 virus by site-directed mutagenesis and subsequent serial passage in ferrets. The genetically modified A/H5N1 virus acquired mutations during passage in ferrets, ultimately becoming airborne transmissible in ferrets. None of the recipient ferrets died after airborne infection with the mutant A/H5N1 viruses. Four amino acid substitutions in the host receptor-binding protein hemagglutinin, and one in the polymerase complex protein basic polymerase 2, were consistently present in airborne-transmitted viruses. The transmissible viruses were sensitive to the antiviral drug oseltamivir and reacted well with antisera raised against H5 influenza vaccine strains. Thus, avian A/H5N1 influenza viruses can acquire the capacity for airborne transmission between mammals without recombination in an intermediate host and therefore constitute a risk for human pandemic influenza.

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          Most cited references 56

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          Universal primer set for the full-length amplification of all influenza A viruses.

           E. Hoffmann,  J Stech,  Y Guan (2001)
          To systematically identify and analyze the 15 HA and 9 NA subtypes of influenza A virus, we need reliable, simple methods that not only characterize partial sequences but analyze the entire influenza A genome. We designed primers based on the fact that the 15 and 21 terminal segment specific nucleotides of the genomic viral RNA are conserved between all influenza A viruses and unique for each segment. The primers designed for each segment contain influenza virus specific nucleotides at their 3'-end and non-influenza virus nucleotides at the 5'-end. With this set of primers, we were able to amplify all eight segments of N1, N2, N4, N5, and N8 subtypes. For N3, N6, N7, and N9 subtypes, the segment specific sequences of the neuraminidase genes are different. Therefore, we optimized the primer design to allow the amplification of those neuraminidase genes as well. The resultant primer set is suitable for all influenza A viruses to generate full-length cDNAs, to subtype viruses, to sequence their DNA, and to construct expression plasmids for reverse genetics systems.
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            Mapping the antigenic and genetic evolution of influenza virus.

            The antigenic evolution of influenza A (H3N2) virus was quantified and visualized from its introduction into humans in 1968 to 2003. Although there was remarkable correspondence between antigenic and genetic evolution, significant differences were observed: Antigenic evolution was more punctuated than genetic evolution, and genetic change sometimes had a disproportionately large antigenic effect. The method readily allows monitoring of antigenic differences among vaccine and circulating strains and thus estimation of the effects of vaccination. Further, this approach offers a route to predicting the relative success of emerging strains, which could be achieved by quantifying the combined effects of population level immune escape and viral fitness on strain evolution.
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              The influenza virus resource at the National Center for Biotechnology Information.

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                Author and article information

                Journal
                10.1126/science.1213362
                22723413

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