West Nile virus is a mosquito-borne flavivirus originally isolated in 1937 from the blood of a febrile woman in the West Nile province of Uganda. The virus is widely distributed in Africa, Europe, Australia, and Asia, and, since 1999, it has spread rapidly throughout the western hemisphere, including the USA, Canada, Mexico, and the Caribbean and into parts of Central and South America. Before 1994, outbreaks of West Nile virus were sporadic and occurred primarily in the Mediterranean region, Africa, and east Europe. Since 1994, outbreaks have occurred with a higher incidence of severe human disease, particularly affecting the nervous system. In North America, the virus has caused meningitis, encephalitis, and poliomyelitis, resulting in significant morbidity and mortality. The goal of this Review is to highlight recent advances in our understanding of West Nile virus virology, ecology, clinical disease, diagnosis, and development of potential vaccines and antiviral therapies.