This study detected and characterized the TevAT1 gene of Trypanosoma evansi isolates from Philippine water buffaloes (Bubalus bubalis). A total of 68 blood samples from Philippine water buffaloes were subjected to DNA extraction and PCR assay was performed using RoTat 1.2 gene to detect T. evansi. Those samples positive for T. evansi subsequently underwent another PCR assay to detect the presence of TevAT1 gene. Trypanosoma evansi was detected in 26.47% (18/68) blood samples in which distributed throughout the main islands of the country (4 from Luzon, 2 from Visayas and 12 from Mindanao). However, only 10 of these samples were positive for TevAT1 gene. Sequence alignment of the TevAT1 gene from local isolates showed no single nucleotide polymorphisms when compared to other strains in various countries. Those T. evansi without the gene of interest could be possibly resistant to some trypanocidal drugs but this needs to be further investigated in-vitro or in-vivo.