The 2014-16 Ebola virus (EBOV) outbreak in West Africa has attracted widespread concern. Rapid and sensitive detection methods are urgently needed for diagnosis and treatment of the disease. Here, we propose a novel method for EBOV detection based on efficient amplification of electroluminescent nanospheres (ENs) coupled with immunomagnetic separation. Uniform ENs are made by embedding abundant amounts of CdSe/ZnS quantum dots (QDs) into copolymer nanospheres through simple ultrasound. Compared to QDs, ENs can enhance electroluminescence (ECL) signals by approximately 85-fold, achieving a signal-to-background ratio high enough for EBOV detection. The introduction of magnetic nanobeads (MBs) can selectively separate targets from complex samples, simplifying the operation process and saving time. The presence of MBs can amplify ECL by approximately 3-fold, improving detection sensitivity. By integration of ENs with MBs, a sensitive electroluminescence biosensor is established for EBOV detection. The linear range is 0.02-30 ng/mL with a detection limit of 5.2 pg/mL. This method provides consistent reproducibility, specificity, and anti-interference ability and is highly promising in clinical diagnosis applications.