In this study, chicken infectious anemia virus (CIAV) was detected from two unvaccinated commercial broiler flocks, showing clinical signs relevant to Chicken infectious anemia (CIA) disease. The clinical signs include subcutaneous hemorrhage, depression, and death. Necropsy findings were pale liver, severe atrophy of bursa of Fabricius and thymus, and discoloration of the bone marrow. DNA was extracted from the liver of infected birds and the VP1 gene was detected by a polymerase chain reaction. The virus strains were detected and genetically characterized in almost the whole VP 1 gene (1338 base pairs). The nucleic acid analysis showed that the two Palestinian strains have 99.9% similarity, and 98.7% similarity to a strain from chicken in Bangladesh (Accession no. AF395114) and 98.5% to a strain from a human fecal sample (Accession no. JQ690762) from China in the same cluster. The lowest similarity (94.7%) was with a strain isolated from chicken in China (Accession no. KU645525). Alignment of the deduced VP1 amino acids showed that the two Palestinian strains shared 100% homology (with one silent mutation encodes Leucine at residue no 97(97L)), classified as high pathogenicity based on glutamine at residue at position 394 (394Q). The highest similarity was with the Bangladesh 99.8% (AAM20899), Brazil 99.5% (APQ44719), and China (AAZ40209) 99.5% strains, while the lowest was with China strain (AFR46599) 96.6%. The study showed valuable information on the molecular characterization of CIAV strains in Palestine for the first time.