Abstract Cystic echinococcosis (CE) is a zoonotic parasitic disease caused by the larval stage; hydatid cyst, of E. granulosus. The disease is usually asymptomatic, and mostly presents by pressure symptoms according to its location. The diagnosis of CE is currently based on imaging techniques, which may not be available in some areas and antibody detection in serum, with several known drawbacks. Conversely, antigen detection has proven its efficacy in serodiagnosis of CE. The study aimed to test the diagnostic efficacy of Nanomagnetic beads-enzyme linked immunosorbent assay (NMB-sandwich ELISA) for diagnosis of CE by detection of E. granulosus AgB in serum and urine samples in comparison to conventional sandwich ELISA. The study included 42 individuals classified into 3 groups; CE infection group (Gp I), other parasitic infections group (Gp II) and healthy control group (Gp III). E. granulosus AgB was prepared from human hydatid cysts and used to produce anti- E. granulosus AgB-IgG-pAbs in rabbit which was used to detect AgB level in serum and urine samples by sandwich ELISA and NMB-sandwich ELISA. Using sandwich ELISA, AgB was detected in serum samples of 88.9% of CE patients with sensitivity and specificity of 88.9% and 91.7%, respectively. In urine samples, sandwich ELISA detected AgB in 83.3% of CE patients and the sensitivity and specificity were 83.3% and 87.5%, respectively. NMB-sandwich ELISA detected AgB in serum samples of 94.4% of CE patients with a sensitivity and specificity of 94.4% and 95.8%, respectively, whereas in urine samples it was detected in 88.9% of CE cases and the sensitivity and specificity were 88.9% and 95.8%, respectively. There was no statistically significant difference (p > 0.05) between the use of serum and urine samples for detection of AgB when examined by either sandwich ELISA or NMB- sandwich ELISA. The best test accuracy was obtained with NMB-sandwich ELISA detecting AgB in serum samples (95.2%), whereas, it showed a slightly lower accuracy of 92.9% when using urine samples. It was concluded that NMB-sandwich ELISA is efficient for detection of AgB in serum samples. Moreover, urine samples might be a good alternative to serum samples for the detection of AgB showing reasonable sensitivity, specificity and accuracy.