High resolution melting analysis as a tool to detect molecular markers of antiviral resistance in influenza A viruses.
作者： Rosaria Arvia , Fabiana Corcioli , Alberta Azzi
DOI: 10.1016/J.JVIROMET.2013.02.016
关键词: High Resolution Melt 、 Genetics 、 Virology 、 Gene 、 Virus 、 Oseltamivir 、 Influenza A virus 、 Sanger sequencing 、 Mutation 、 Pyrosequencing 、 Biology
摘要: Abstract A real-time PCR followed by high resolution melting analysis (HRMA) was developed, for rapid detection of antiviral resistance markers in influenza viruses, both H1N1 and H3N2 subtypes. The targets these assays were the nucleotide substitution G806A (S31N mutation) M gene as marker to adamatanes viruses A(H3N2), A356T (E119V N2 A(H3N2) C823T (H274Y N1 pandemic A(H1N1) 2009 virus oseltamivir resistance. First, designed primers overall protocol HRMA validated using already characterized viral isolates either containing or lacking changes at tested codons. Then, used search 75 clinical samples, positives, analyzed previously pyrosequencing Sanger sequencing, adamantane-derivatives 57 positive samples. results agreement with those obtained sequencing. As regards isolates, revealed a widespread adamantanes 89.5% G806A, while 3% resistant (A356T change). HRMA, applied drugs against confirmed be procedure flexible, low cost time-saving, suitable application epidemiological surveys settings diagnostic purposes.
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