In order to isolate Neospora caninum in cell culture (Vero cell) and diagnosis the strains by using Real Time Polymerase Chain Reaction by using three different primers pairs(NC-5,Nc1 and BPA4) from the tissue of  25 aborted fetus, 14 new borne calves and 39 cattle which suffer from abortion in different stages of pregnancy, the study showed from culturing of tissue sample extract in tissue culture, Cyto Pathic Effect this effect manifested by Vacule, Rupture in cell and presence of tachyzoites, and this effect variance in density between samples and passages, the first CPE appear in 3^rd passage of aborted fetus heart, in 4^th passage of calves brain and heart, and cow placenta. When application the PCR technique in DNA template which extracted from infected cell culture ,showed presence increased fluorescent curve which indicate positive sample, the highest percentage of infection in cell culture which infected with brain and skeletal muscle of Aborted fetus and Heart, skeletal muscle of calves, and  blood sample of cow, the highest percentage in 2^nd trimester aborted fetus, in calves which suffer from recumbence, and in cow which aborted in 2^nd trimester. The high percentage of NC-5 strain in blood sample of cow, brain of calves, brain and skeletal muscle of aborted fetus, Nc1 strain not appear in blood sample of cow, calves and brain of calves in additional of brain and skeletal muscle of aborted fetus, while highest percentage of BPA4 strain appear in skeletal muscle of calves and aborted fetus and blood of cow