West Nile virus (WNV) infection is an emerging zoonosis that threatens global public health. In this study, a total of 95 bird serum samples from 14 species and 341 horse serum samples were collected from 2008 to 2010 in Shanghai, China. All serum samples were screened initially for WNV-reactive antibodies using a competitive ELISA. The positive samples detected by ELISA were further confirmed using a plaque-reduction neutralization test (PRNT) for WNV and its most closely related flaviviruses in the area to avoid false positives due to cross-reactivity. Five (5·3%) of the bird serum samples and none (0·0%) of the horse serum samples tested positive for WNV antibodies. The findings strongly suggest that some of the birds, specifically the resident birds in China, had been exposed to WNV.
