Rapid detection of influenza A pandemic (H1N1) 2009 virus neuraminidase resistance mutation H275Y by real-time reverse transcriptase PCR

Abstract

The emergence of oseltamivir-resistant influenza A pandemic (H1N1) 2009 virus highlights the need for rapid oseltamivir resistance screening. We report the development and validation of high-throughput real-time reverse transcriptase PCR assays for the detection of the H275Y substitution in the neuraminidase 1 gene that can be accomplished in 3 to 4 h.

FIG. 1.

Linear limits of detection of the H275Y (A) and H275 (B) qRT-PCR assays. Serial (10-fold) dilutions of influenza A pandemic (H1N1) 2009 virus resistant (A) or sensitive (B) to oseltamivir were prepared and tested in qRT-PCR assays. C_t values were obtained for each dilution and are plotted against the TCID₅₀ value.