牛奶过敏原PCR检测方法的建立

牛奶是主要的食品过敏原之一,其中β-乳球蛋白是引起牛奶过敏的主要成分。基于牛奶β-乳球蛋白基因序列设计一对PCR引物,经过反应条件的优化,建立了适合检测牛奶β-乳球蛋白过敏原的PCR方法。实验结果表明,当PCR反应体系中模板DNA量为25ng,引物浓度为4mmol/L时,检测效果最佳。所建立的方法不仅可用于不同保质期、不同来源牛奶β-乳球蛋白过敏原的测定,亦可用于牛、羊奶不同比例混合物中β-乳球蛋白过敏原的检测。

PCR method for the detection of allergen in milk

Milk is considered to be a kind of important allergen,and β - lactoglobulin is one major component of allergens in milk. A pair of primers was designed based on the gene sequence of β - lactoglobulin in milk and PCR reaction system was optimized for detecting β - lactoglobulin allergen. The results showed that best results could be obtained by combining 25ng template DNA with 4mmol/L primers used for PCR analysis. The established method can not only be successfully applied in detecting β - lactoglobulin allergen in milk from different shelf lives,and resources,but also in detecting the allergen in the mixture at different ratios of cow milk and goat milk.