Oseltamivir activity against avian influenza H9N2 strain with different point mutations in their neuraminidase

Abstract

The present study has two aims: to optimize the antiviral activity of oseltamivir in chicken embryos against an avian influenza-H9N2 strain (P0) and to apply the optimized protocol for studying the drug susceptibility of 4 H9N2 mutants (M1, M2, M3, and M4). As for the first aim, oseltamivir antiviral activity was monitored upon its delivery into 9-day-old chicken embryo at a concentration of 0.27 mg/100 μl, against 7 doses of the P0 strain, ranging between 1.2 × 10-5 and 2.0 Hemagglutination (HA) units. Oseltamivir showed its highest efficacy in reduction of viral propagation (95% reduction in HA titer) (P < 0.05), when the inoculum level contained a minimum HA units of 1.2 × 10-5. For the second aim of this study, the application of the 1.2 × 10-5 HA units of the virus in inocula for the evaluation of oseltamivir‑antiviral effect against the 4 H9N2 mutants revealed an emergence of a resistant mutant (M1), associated with 2 adjacent point mutations in its neuraminidase (N) amino acid (aa) sequence at positions 46 and 47. The other 3 mutants maintained a variable sensitivity to oseltamivir, resulting in the following reduction in HA titers: M2 (82.9%), M3 (61.5%), and M4 (100.0%). How the point mutations of the neuraminidase sequences affected the susceptibility of H9N2 virus to oseltamivir is still to be determined and deserve further investigations. © 2015, Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise “G.Caporale”. All rights reserved.