Oseltamivir is the most effective antiviral drug used for the treatment and prevention of influenza A infections. Neuraminidase is the principal target for treating patients with H5N1 infection. Until recently, only a low prevalence of neuraminidase inhibitors (NAIs) resistance (<1 %) had been detected in circulating viruses. However, there have been reports of significant numbers of influenza A (H5N1) strains with a H274Y neuraminidase mutation that was highly resistant to the NAI, oseltamivir. In this study, we used molecular docking and molecular dynamics (MD) approach to characterize the effect of H274Y mutation in drug-target interactions. Docking results suggest that oseltamivir was found to adopt the most promising conformations to the wild type NA (WT) by identifying the guanidinium side chain of R-156 and R-152 as a prospective partner for making polar contacts as compared to the mutant type NA. The MD results showed that the average atom, especially atoms of the wild type NA-oseltamivir complex, movements were small, displayed fast convergence of energy and charges in geometry. This highlights the stable binding of the oseltamivir with wild type NA as compared to mutant type NA. Overall, our study may be helpful for the rational design of more powerful, selective, and more robust NAI against drug-resistant H274Y mutation. © 2013 Springer Science+Business Media New York.