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Objective To establish a high-throughput sequencing method for detecting cereal allergen species based on next-generation sequencing technology. Methods Taking Hordeum vulgare, Triticum aestivum, Fagopyrum esculentum and Fagopyrum tataricum as research objects, next-generation sequencing technology was used for the high-throughput identification of cereal allergen samples with different proportions and mixing methods and commercially available samples were collected to verify the method’s applicability. Results In different proportions of simulated samples, Hordeum vulgare, Triticum aestivum, Fagopyrum esculentum and Fagopyrum tataricum were detected, the limit of detection was as low as 0.1%, but the measured relative abundance of species and the actual amount of different additives; in different mixing methods but the same proportion of simulated samples, cereal allergens had been detected; the relative abundance of DNA mixing and mixing of raw materials and raw material mixing methods were different from the actual amount of species, and raw material mixing methods of species relative abundance closer to the exact amount of additives; commercial samples contained varying degrees of grain allergens, while a small number of samples with the ingredient list or label. Conclusion The detection method established in this study enables specific and simultaneous identification of 4 cereal allergens (Triticum aestivum, Hordeum vulgare, Fagopyrum esculentum and Fagopyrum tataricum). By conducting relative quantitative analysis of species abundance, the method can effectively determine the content of cereal allergens in samples.
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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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