A sandwich ELISA method based on immunomagnetic beads for the rapid enrichment and detection of Tilletia foetida teliospores in wheat and flour samples

Common bunt is a major disease of wheat worldwide that reduces crop yields and grain quality. Rapid and sensitive quantitative detection methods are required to diagnose and monitor this disease in wheat management programs and to ensure seed quality. In this study, an immunomagnetic beads-based enzyme-linked immunosorbent assay (IMBs-ELISA) was developed for the detection of Tilletia foetida teliospores in wheat and flour. An anti-T. foetida teliospores polyclonal antibody bound to immunomagnetic beads was used as the capture probe, and a polyclonal antibody labeled with horseradish peroxidase was used as the detector probe. The capture and detection conditions for the target spores were optimized to achieve the best determination results. Under optimal conditions, the proposed method took less than 2 h to complete. Its limit of detection was 300 teliospores per gram. The critical reaction in this IMBs-ELISA occurred on magnetic beads. This not only simplified the traditional ELISA process, but also shortened the detection time. This study has expanded the application of the IMBs-ELISA method to fungal spore detection. This method has potential applications in agriculture and seed management.