Abstract: C6/36 culture isolation test, indirect immunofluorescence assay, and reverse transcription PCR and sequencing of the PCR products were carried out to isolate and identify the West Nile virus （WNV） from experimentally infected Culex tritaeniorhynchus and Leghorn chicken. The cell pathological effects such as cell fusion and cavitation were observed in C6/36 inoculated experimentally infected samples. The antigen of WNV was detected by indirect immunofluorescence assay. Three specific PCR bands with expected size (408 bp, 498 bp, and 559 bp, respectively) were found, and the sequences of these PCR products were concordant with those of WNV Chin-01 strain. Hence，it was confirmed that the WNV from experimentally infected Cx. Tritaeniorhynchus and Leghorn chicken was same as WNV Chin-01 strain used in our experiments.

Key words: Culex tritaeniorhynchus, Leghorn chicken, West Nile virus, isolation, identification