Monitoring the susceptibility to oseltamivir of Influenza A(H1N1) 2009 virus by nested-PCR and pyrosequencing during the pandemic and in the season 2010-2011.
For the early detection of the H275Y mutation as a marker of oseltamivir resistance in A(H1N1) pandemic strains, a sensitive and specific pyrosequencing assay was developed. This assay analyses a region 99nts long, encompassing the H275Y site, amplified by a nested PCR. Seventy-five respiratory specimens, obtained from 62 patients during the pandemic and in the 2010-2011 influenza season, in Tuscany, were tested. Resistant strains were demonstrated in 10 patients. In three other patients, resistant and sensitive variants were found. This pyrosequencing assay may be a useful method for monitoring the spread of resistant influenza H1N1 2009 strains.
Public Health Department, University of Florence, Italy.
Rosaria
R
Arvia
Fabiana
F
Corcioli
Lisa
L
Simi
Claudio
C
Orlando
Riccardo
R
De Santis
Marzia
M
Facchini
Isabella
I
Donatelli
Alberta
A
Azzi
D000293:Adolescent; D000328:Adult; D019943:Amino Acid Substitution; D000998:Antiviral Agents / Q000494:pharmacology*; D002648:Child; D002675:Child, Preschool; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D053118:Influenza A Virus, H1N1 Subtype / Q000187:drug effects*; D007251:Influenza, Human / Q000821:virology; D007558:Italy; D008826:Microbial Sensitivity Tests / Q000379:methods; D020125:Mutation, Missense; D053139:Oseltamivir / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000737:chemistry* / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*
D000998:Antiviral Agents; D012367:RNA, Viral; D053139:Oseltamivir