基于高通量测序技术的谷物过敏原识别研究

Identification of cereal allergens based on high-throughput sequencing technology

中文关键词:  谷物过敏原  麸质过敏原  无麸质过敏原  高通量检测

英文关键词:cereal allergens  gluten allergens  gluten-free allergens  high-throughput detection technology

中文摘要:

      目的 建立基于下一代测序技术的谷物过敏原物种高通量检测方法。方法 以大麦、小麦、甜荞、苦荞为研究对象, 针对不同比例、不同混合方式的谷物过敏原样品, 采用下一代测序技术进行高通量鉴别, 并收集市售样品对方法适用性进行验证。结果 在不同比例的模拟样品中, 小麦、大麦、甜荞、苦荞均被检出, 检出限低至0.1%, 但所测得的物种相对丰度与实际添加量不同; 在不同混合方式但比例相同的模拟样品中, 谷物类过敏原均被检出, DNA混合与原料混合方式测得的物种相对丰度均与实际添加量不同, 且原料混合方式所得的物种相对丰度更接近实际添加量; 市售样品中含有不同程度的谷物过敏原, 同时存在少部分样品与配料表或标签不符的情况。结论 本研究建立的检测方法可实现小麦、大麦、甜荞及苦荞4类谷物类过敏原的特异性同步鉴别, 并通过物种丰度的相对定量分析, 可以有效判别样品中谷物类过敏原物种成分的含量高低。

英文摘要:

      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.