This report describes the development of a sensitive and broadly specific indirect competitive enzyme-linked immunosorbent assay (icELISA) and a gold nanoparticle-based immunochromatographic strip (GNP-ICS) assay for the detection of benzo[a]pyrene (B[a]P), using an anti-B[a]P monoclonal antibody (mAb). A broad-specific anti-B[a]P mAb (4E8) was raised from two types of haptens, with half maximal inhibitory concentrations and limits of detection (LOD) values of 2.51 and 0.54 ng mL-1, respectively. In addition, the cross-reactivity was up to 390% with structurally related compounds. The GNP-ICS assay based on a GNP-labeled mAb showed broad specificity in the detection of B[a]P and its analogues, with cut-off and visual LOD values of 100 and 10 ng mL-1, respectively. Furthermore, the recoveries from the developed icELISA and GNP-ICS assay in edible oil samples spiked with B[a]P were validated by high-performance liquid chromatography-fluorescence detection. The results revealed that the icELISA could reliably detect B[a]P in edible oils.