期刊
ANTIVIRAL RESEARCH
卷 81, 期 1, 页码 16-24出版社
ELSEVIER
DOI: 10.1016/j.antiviral.2008.08.008
关键词
Antiviral; Influenza A virus; Oseltamivir; Zanamivir; Pyrosequencing
资金
- U.S. State Laboratories, the National Influenza Centers
- Influenza Surveillance Network and the WHO Collaborating Centers
- Oak Ridge Institute for Science and Education (ORISE)
Pyrosequencing has been successfully used to monitor resistance in influenza A viruses to the first class of anti-influenza drugs, M2 blockers (adamantanes). In contrast to M2 blockers, resistance to neuraminidase (NA) inhibitors (NAIs) is subtype- and drug-specific. Here, we designed a pyrosequencing assay for detection of the most commonly reported mutations associated with resistance to NAIs, a newer class of anti-influenza drugs. These common mutations occur at residues: H274 (N1), E119 (N2), R292 (N2), and N294 (N2) in seasonal influenza A viruses. Additionally, we designed primers to detect substitutions at D151 in NAs of N1 and N2 subtypes. This assay allows detection of mutations associated with resistance not only in grown viruses but also in clinical specimens, thus reducing the time needed for testing and providing an advantage for disease outbreak investigation and management. The pyrosequencing approach also allows the detection of mixed populations of virus variants at positions of interest. Analysis of viruses in the original clinical specimens reduces the potential for introducing genetic variance in the virus population due to selection by cell culture. Our results showed that, in at least one instance, a D151 E change seen in N1NA after virus propagation in cell culture was not detected in the original clinical specimen. Although the pyrosequencing assay allows high throughput screening for established genetic markers of antiviral resistance, it is not a replacement for the NA inhibition assays due to insufficient knowledge of the molecular mechanisms of the NAI-resistance. Published by Elsevier B.V.
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