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Objective To establish a rapid visual detection method for Aspergillus niger in Zea mays L. by loop-mediated isothermal amplification (LAMP) technology. Methods Based on LAMP technology, specific primers targeting the key ochratoxin A (OTA) biosynthesis gene OTAhal were designed. Through screening of the primers and optimization of conditions such as primer sequence, primer ratio, reaction temperature and visual dye, the optimal detection conditions were determined. Using the total DNA of Aspergillus niger and artificially contaminated corn samples as templates, the sensitivity of the detection method and the feasibility of its practical application were validated. Results The optimal primer ratio was 8:4:1, the optimal reaction temperature was 64 °C, and the method could complete the amplification of target DNA within 30 minutes. The color of the positive sample changed from violet to blue-purple. This method could detect DNA concentrations as low as 6.87×10⁻3 ng/μL. When applied to actual Zea mays L. sample testing, it achieved a detection limit as low as 10¹ spores/mL. Conclusion This method is simple to operate, highly sensitive, and does not require sophisticated instruments. It can be applied to the detection of OTA in actual Zea mays L. samples, indicating broad application prospects.
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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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