常规PCR法同时检测食品中的小麦、大麦和黑麦致敏原

Simultaneous determination of wheat, barley and rye allergen in food by conventional PCR

中文关键词:  小麦  大麦  黑麦  谷蛋白  PCR  乳糜泻

英文关键词:wheat  barley  rye  gluten  PCR  coeliac disease

中文摘要:

      目的 建立一种常规PCR法同时检测食品中的小麦、大麦和黑麦致敏原(WBR155bp)。方法 采用ClustalW软件分析小麦ω-醇溶谷蛋白、大麦醇溶谷蛋白和黑麦ω-黑麦碱的核苷酸编码序列的同源性, 进而确定同源区域, 用Primer Premier5.0基于同源区域设计引物。选取不同种属的植物考查所建立PCR方法的特异性。将小麦粉, 大麦粉、黑麦粉与米粉混合制备谷物含量梯度为100%、10%、1.0%、0.1%、0.01%的样品, 考查WBR155 bp的检出限。结果 扩增产物片段<200 bp, 可以检测到高度降解的DNA片段。引物WBR155bp对小麦、大麦和黑麦的检出限分别为1.0%, 0.01%和0.1%。结论 该方法具有良好特异性, 可用于食品中的小麦、大麦和黑麦致敏原的日常检测。

英文摘要:

      Objective To establish a method for simultaneous determination of wheat, barley and rye allergen (WBR155bp) in food by conventional PCR method. Methods ClustalW software was used to analyze the homology of the nucleotide coding sequences of wheat ω-gliadin, hordein and ω-secalin, and then to determine the homologous region. Primer Premier 5.0 was used to design primers based on homologous regions. The specificity of the PCR method was tested by selecting plants of different species. The grain content gradient of wheat flour, barley flour, rye flour and rice flour was 100%, 10%, 1.0%, 0.1%, and 0.01%, respectively. The detection limit of WBR155 bp was examined. Results Amplified product fragment was <200 bp, and highly degradable DNA fragments could be detected. The detection limits of primer WBR155bp on wheat, barley and rye were 1.0%, 0.01% and 0.1%, respectively. Conclusion This method has good specificity and can be used for routine detection of wheat, barley and rye allergens in food.