An immunosensor for the rapid detection of 1,3-dinitrobenzene was developed based on an evanescent wave all-fiber biosensing platform with the detection limits of 0.054 mg x L(-1), and the detection cycle was less than 10 min. Hapten-carrier conjugates NB-OVA were synthesized by mixing 4-nitrohippuric acid and OVA activated by EDC, and then the conjugates were immobilized onto the silane layer on the probe with a heterobifunctional crosslinker. The probe modified had good robustness and regeneration performance, which allowed the performance of more than 100 assay cycles without significant loss of reactivity. Several water samples of different origins were measured with less than 4.5% -10.0% deviation of the detection and the recovery rate of 1,3-dinitrobenzene was between 80% and 120%, which proved the system's precision and accuracy and negligible matrix effects. This immunosensor shows great potential in rapid detection of 1,3-dinitrobenzene in practical waters.