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Objective To develop specific antibodies against Fusarium graminearum and establish a double-antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) for detecting the contamination of Fusarium graminearum in Triticum aestivum. Methods The fungal lysate antigen of Fusarium graminearum was prepared by physical grinding. Monoclonal antibody (mAb) against Fusarium graminearum were developed using hybridoma technology. The performance parameters of mAbs, such as specificity and affinity, were measured. The optimal combination conditions of the prepared mAbs and polyclonal antibody (pAb) were explored using the checkerboard method, and then the DAS-ELISA method was established. The sensitivity and repeatability of the method were evaluated, and the method was applied to the detection of naturally infected Triticum aestivum samples. Results The mAb 1C4F3 and pAb FG2801 against Fusarium graminearum were successfully prepared. The mAb showed no cross-reaction with other fungi, indicating strong specificity. The affinity constant was 1.26×10⁷ L/mol, suggesting high affinity. A DAS-ELISA method for Fusarium graminearum was established, with a limit of detection of 1.223 μg/mg and a standard curve range of 1.56-100.00 μg/mg. The reliability of the method was verified by detecting 96 naturally infected samples collected from major Triticum aestivum-producing areas in China. A significant positive correlation was found between the content of Fusarium graminearum and the content of deoxynivalenol (DON) in the samples (r2=0.8897). Therefore, the DAS-ELISA could not only be used for the rapid detection of Fusarium graminearum but also for evaluating the DON contamination in the samples. Conclusion The DAS-ELISA developes in this study enables rapid and quantitative determination of Fusarium graminearum in Triticum aestivum. Simultaneously, it allows for a swift assessment of the DON contamination level in samples. Characterized by high accuracy and reliability, this method holds considerable application potential in monitoring Fusarium graminearum contamination in Triticum aestivum crops both in the field and post-harvest.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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