Objective. To investigate the presence of WNV in birds, horses and humans in northeast Mexico. Material and Methods. Serum samples from 33 birds, 24 horses and 237 humans were screened by ELISA for Anti-WNV antibodies. Human serum samples were also screened for WNV RNA using an RT-PCR assay. Results. Positive sera were found in three birds and 15 horses. Forty percent of the human serum samples were positive for IgG antibodies and 0% for IgM antibodies and viral RNA. Conclusions. The results of this study show that WNV is present in northeast Mexico and it is a new emergent infectious agent that represents a challenge for research and prevention programs in Mexico.