Development and implementation of influenza a virus subtyping and detection of genotypic resistance to neuraminidase inhibitors
Identifieur interne : 000F60 ( Main/Exploration ); précédent : 000F59; suivant : 000F61Development and implementation of influenza a virus subtyping and detection of genotypic resistance to neuraminidase inhibitors
Auteurs : Guillermo Ruiz-Carrascoso [Espagne] ; Inmaculada Casas [Espagne] ; Francisco Pozo [Espagne] ; Carmen Pérez-González [Espagne] ; Jordi Reina [Espagne] ; Pilar Pérez-Bre A [Espagne]Source :
- Journal of Medical Virology [ 0146-6615 ] ; 2010-05.
English descriptors
- Teeft :
- Adamantane resistance, Agarose, Amersham pharmacia, Amino, Amino acid sequences, Amino acids, Analytical sensitivity, Animal origin, Antigenic, Antiviral, Antiviral resistance, Antiviral susceptibility, Assay, Avian, Cell culture, Clinical microbiology, Clinical practice, Clinical samples, Clinical specimens, Consensus sequence, Consensus sequences, Diagnostic purposes, Different host species, Embryonated chicken eggs, Enzyme mixture, Epidemiological week, Ethidium bromide, Genbank accession, Genbank numbers, Genetic characterization, Genetic clusters, Govorkova, Gran canaria, Gubareva, Hemagglutinin, High incidence, High rates, High sensitivity, Human metapneumovirus, Human origin, Human strains, Human virus, Immunocompromised child, Inhibitor, Magattract virus mini, Molecular size marker, Multiple alignments, Multiplex, Mutation, Nais, Nasopharyngeal aspirates, National center, Neuraminidase, Neuraminidase inhibitor resistance, Neuraminidase inhibitors, Northern hemisphere, Nucleic acids, Nucleotide, Oseltamivir, Oseltamivir resistance, Pandemic, Primer, Primer sets, Prototype strains, Reaction mixture, Reaction volume, Reference laboratories, Resistance markers, Resistant viruses, Respiratory syncytial viruses, Respiratory viruses laboratory, Second round, Sensitive viruses, Sequencing, Serial dilutions, Similar approach, Simultaneous detection, Spanish ministerio, Spanish surveillance network, Step conditions, Substitution, Subtype, Subtypes, Subtyping, Taqman probes, Universal primers, University hospital, Useful tool, Vaccine, Vaccine strains, Viral, Virol, Virus, Virus resistance.
Abstract
Influenza virus hemagglutinin and neuraminidase, surface glycoproteins with an essential role in viral pathogenesis, are important antigen determinants and essential markers for epidemiological surveillance. Neuraminidase is also a suitable target for designing antiviral drugs. The introduction into clinical practice of neuraminidase inhibitors and the development of random point mutations have increased the emergence of drug‐resistant viruses. A universal RT nested PCR‐based system has been developed for subtyping H1, H3, N1 and N2, in influenza A viruses of human or animal origin. The subsequent sequencing and analysis of the hemagglutinin and neuraminidase templates reveal antigenic and receptor binding changes in the HA1 subunit and mutations of clinical relevance concerning resistance to neuraminidase inhibitors. The specificity and sensitivity of the method were evaluated using 113 influenza A isolates, 105 influenza A positive respiratory samples obtained from patients and 29 prototype strains of both human and animal origin. The resulting analytical sensitivity of the subtyping techniques is one to at least 100 molecules of cloned DNA product in a final reaction volume of 50 µl. In the course of implementing the method, two H1N1 isolates with the H274Y mutation in the neuraminidase segment have been detected and their molecular features analyzed. The emergence of influenza virus resistance makes the neuraminidase genetic characterization and surveillance activities to detect antiviral resistance necessary. J. Med. Virol. 82: 843–853, 2010. © 2010 Wiley‐Liss, Inc.
Url:
DOI: 10.1002/jmv.21692
Affiliations:
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Neuraminidase is also a suitable target for designing antiviral drugs. The introduction into clinical practice of neuraminidase inhibitors and the development of random point mutations have increased the emergence of drug‐resistant viruses. A universal RT nested PCR‐based system has been developed for subtyping H1, H3, N1 and N2, in influenza A viruses of human or animal origin. The subsequent sequencing and analysis of the hemagglutinin and neuraminidase templates reveal antigenic and receptor binding changes in the HA1 subunit and mutations of clinical relevance concerning resistance to neuraminidase inhibitors. The specificity and sensitivity of the method were evaluated using 113 influenza A isolates, 105 influenza A positive respiratory samples obtained from patients and 29 prototype strains of both human and animal origin. The resulting analytical sensitivity of the subtyping techniques is one to at least 100 molecules of cloned DNA product in a final reaction volume of 50 µl. In the course of implementing the method, two H1N1 isolates with the H274Y mutation in the neuraminidase segment have been detected and their molecular features analyzed. The emergence of influenza virus resistance makes the neuraminidase genetic characterization and surveillance activities to detect antiviral resistance necessary. J. Med. Virol. 82: 843–853, 2010. © 2010 Wiley‐Liss, Inc.</div></front></TEI><affiliations><list><country><li>Espagne</li></country><region><li>Communauté de Madrid</li></region><settlement><li>Madrid</li></settlement></list><tree><country name="Espagne"><region name="Communauté de Madrid"><name sortKey="Ruiz Arrascoso, Guillermo" sort="Ruiz Arrascoso, Guillermo" uniqKey="Ruiz Arrascoso G" first="Guillermo" last="Ruiz-Carrascoso">Guillermo Ruiz-Carrascoso</name></region><name sortKey="Casas, Inmaculada" sort="Casas, Inmaculada" uniqKey="Casas I" first="Inmaculada" last="Casas">Inmaculada Casas</name><name sortKey="Perez Onzalez, Carmen" sort="Perez Onzalez, Carmen" uniqKey="Perez Onzalez C" first="Carmen" last="Pérez-González">Carmen Pérez-González</name><name sortKey="Perez Re A, Pilar" sort="Perez Re A, Pilar" uniqKey="Perez Re A P" first="Pilar" last="Pérez-Bre A">Pilar Pérez-Bre A</name><name sortKey="Pozo, Francisco" sort="Pozo, Francisco" uniqKey="Pozo F" first="Francisco" last="Pozo">Francisco Pozo</name><name sortKey="Reina, Jordi" sort="Reina, Jordi" uniqKey="Reina J" first="Jordi" last="Reina">Jordi Reina</name><name sortKey="Ruiz Arrascoso, Guillermo" sort="Ruiz Arrascoso, Guillermo" uniqKey="Ruiz Arrascoso G" first="Guillermo" last="Ruiz-Carrascoso">Guillermo Ruiz-Carrascoso</name><name sortKey="Ruiz Arrascoso, Guillermo" sort="Ruiz Arrascoso, Guillermo" uniqKey="Ruiz Arrascoso G" first="Guillermo" last="Ruiz-Carrascoso">Guillermo Ruiz-Carrascoso</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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