Echinococcus granulosus is a cosmopolitan parasite causing cystic echinococcosis in humans and livestock. Hydatid disease is still a serious public health and economic problem. Is characterized by high intra- specific variability genotypes (G1- G10), the most common strain is sheep strain (G1) which is mostly responsible of human Echinococcosis. The aim of this study was to prove molecular characterization of Echinococcus granulosus (G1) isolates which were obtained from human and livestock (sheep, cow and goat) by using polymerase chain reaction (PCR) in Kirkuk province of Iraq. A total of (40)  isolates were obtained from patients after surgical operation and from Kirkuk- slaughter house, DNA samples were extracted from germinal layer of (7) human, (11) sheep, (17) cow and (5) goat. The specific primer for sheep strain G1 was provided for molecular characterization studies. PCR resulted in a specific product of 400 bp. All isolates genotyped were clearly demonstrated G1 strain using nicotinamide adenine dinucleotide dehydrogenase subunit 1 (nad1) gene. This is the first molecular study report genotyping of Echinococcus isolates in Kirkuk province, Iraq.