Broad-spectrum antibodies can effectively recognize substances with similar structures and have broad application prospects in field rapid detection. In this study, broad-spectrum antibodies (Abs) against organophosphorus pesticides (OPs) were used as sensitive recognition elements, which could effectively recognize most OPs. Gold nanoparticles (AuNPs) have good biocompatibility. It combined with Abs to form a gold-labeled probe (AuNPs-Abs), which enhances the effective binding of antibodies to nanomaterials. Prussian blue (PB) was added to electrodeposition solution to enhance the conductivity, resulting in superior electrochemical performance. The AuNP-Abs-PB composite film was prepared by electrodeposition on the electrode surface to improve the anti-interference ability and stability of the immunosensor. Under the optimal experimental conditions, the immunosensor had a wide detection range (IC-IC: 1.82 × 10-3.29 × 10 ng/mL) and high sensitivity. Most importantly, it was simple to be prepared and could be used to detect multiple OPs.