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Detection of influenza A H1N1 and H3N2 mutations conferring resistance to oseltamivir using rolling circle amplification.

Abstract：

:In the event of an influenza pandemic, the use of oseltamivir (OTV) will undoubtedly increase and therefore it is more likely that OTV-resistant influenza strains will also arise. OTV-resistance genotyping using sequence-based testing on viruses isolated in cell culture is time consuming and less likely to detect the low-level presence of drug-resistant virus populations. We have developed a novel rolling circle amplification (RCA) method to achieve the sensitive detection of OTV-resistant viruses from clinical specimens. Using artificially created templates, RCA could detect the presence of OTV-resistant mutations (N2: 119V, 292K, N1: 274Y) even if the population carrying the mutations was <1% of the total. By applying RCA to clinical samples, we identified the emergence of the 274Y mutation in one OTV-treated patient, as well as in seven individuals who were treatment-naïve (confirming community transmission of 274Y-containing resistant influenza A H1N1). These results were further confirmed by neuraminidase region sequencing. In conclusion, RCA technology can provide rapid (<24 h), high-throughput diagnosis of OTV resistance mutations with a high specificity and sensitivity.

journal_name

Antiviral Res

journal_title

Antiviral research

authors

Steain MC,Dwyer DE,Hurt AC,Kol C,Saksena NK,Cunningham AL,Wang B

doi

10.1016/j.antiviral.2009.09.010

subject

Has Abstract

pub_date

2009-12-01 00:00:00

pages

242-8

issue

3

eissn

0166-3542

issn

1872-9096

pii

S0166-3542(09)00463-X

journal_volume

84

pub_type

杂志文章

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