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A universal microchip was developed for genotyping Influenza A viruses. It contains two sets of oligonucleotide probes allowing viruses to be classified by the subtypes of hemagglutinin H1–H13, H15, H16 and neuraminidase N1–N9. Additional sets of probes are used to detect H1N1 swine influenza viruses. Selection of probes was done in two steps. Initially, amino acid sequences specific to each subtype were identified, and then the most specific and representative oligonucleotide probes were selected. Overall, between 19 and 24 probes were used to identify each subtype of hemagglutinin HA and neuraminidase NA. Genotyping included preparation of fluorescently labeled PCR amplicons of influenza virus cDNA and their hybridization to microarrays of specific oligonucleotide probes. Out of 40 samples tested, 36 unambiguously identified HA and NA subtypes of Influenza A virus.



Autor: Vladimir A. Ryabinin, Elena V. Kostina, Galiya A. Maksakova, Alexander A. Neverov, Konstantin M. Chumakov , Alexander N. Sinyakov

Fuente: http://plos.srce.hr/



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