A simple and novel immunochromatographic assay (ICA) was developed for specific and sensitive determination of ochratoxin A (OTA) employing microorganism (MO) as reducers and carriers. In this work, microorganism loaded gold nanoparticles ([email protected]) composites were synthesized by reacting Yeast/Lactobacillus (LAB) with HAuCl4 via biosorption and subsequent bioreduction. The synthesis conditions (pH and reaction time) and main parameters (morphology, diameter and functional groups) of [email protected] were investigated. Owing to the large sizes (about 2 μm) and high adsorption abilities of [email protected], the few antibodies were labeled, resulting a lower detection limit for ICA. After optimization, the linear detection of OTA using [email protected] immunoassay ranged from 0.1 to 1.6 ng/mL, and the visual limit of detection (vLOD) was 0.1 ng/mL, which was nearly 8-fold lower than traditional AuNPs-ICA. Meanwhile, the proposed strategy also possessed high specificity for OTA against other mycotoxins. The applicability of the platform was successfully assessed through the detection of OTA in rice, corn, ginger and green bean samples. The results indicated that these creative sensing elements are promising alternatives and bring an innovative inspiration for biological detections.