A total of 330 camels (Camelus dromedarius) of both sexes and with different ages, at different localities in Sharkia Governorate, Egypt, were examined clinically from April 2013 to March 2015. Some adult camels showed emaciation, edema of legs and abdomen which were suspected to be infected by Trypanosoma species. This study aimed to determine the seroprevalence of trypanosomiasis using indirect ELISA test and the results showed that 174 (52.7%) were infected. The infection rate was higher in males (54.8%) than females (42.2%), also, in camels more than 2-years old and in summer and spring seasons. Thirty blood samples from the examined animals by ELISA test (15 positive and 15 negative) were subjected to PCR. The results revealed that 18 samples were positive for T. evansi by PCR. Two positive samples for RoTat1.2 VSG encoding gene were chosen for DNA sequence analysis, one of them was obtained from newly imported camel in a camel farm at Belbais city and the other one belonged to a camel from one of Belbais villages at Sharkia Governorate. Nucleotide sequence alignment of RoTat1.2 VSG gene variants from Egyptian T. evansi showed some heterogeneity with other T. evansi isolates from Egypt. In conclusion, PCR technique is more sensitive and specific than ELISA for the diagnosis of T. evansi infection in camels.