West Nile virus (WNV; genus Flavivirus; family Flaviviridae) is the aetiological agent of an emerging, mosquito-borne disease with great impact on human and animal health. Over the past 15 years, WNV has been responsible for large epidemics mainly in North America but also in Europe, where lineage 1 and more recently lineage 2 strains have caused an upsurge in the number of outbreaks with increased human infection and higher virulence for certain wild bird species. This study aimed to compare the course of infection of the lineage 1 WNV strains Israel/98 and Italy/08 and the lineage 2 strain Austria/08 in the red-legged partridge (Alectoris rufa), a gallinaceous bird indigenous to the Iberian Peninsula and widely distributed in Southern and Western Europe. After experimental inoculation, clinical and analytic parameters (viraemia, viral load, antibodies) were examined over a period of 15 days. All inoculated birds became viremic and showed clinical disease, with a morbidity rate of 100% and mortality rates between 22.2 and 55.5% depending on the virus strain. The red-legged partridge demonstrated to be a competent host for transmission of the three investigated WNV isolates with the highest competence index observed for the Italian strain. Likewise, this strain was the most pathogenic causing the highest viral loads in blood, organs, feathers and oral and cloacal secretions. These experimental results indicate that the red-legged partridge is highly susceptible to the infection with lineage 1 and 2 WNV strains and that this species may act as an amplifying host for both WNV lineages.