食品中牛奶过敏原成分LAMP检测方法的建立

Establishment of loop-mediated isothermal amplification method for detection of milk allergen in foods

中文关键词:  牛奶  过敏原  环介导等温扩增(LAMP)  实时荧光PCR

英文关键词:milk  allergen  loop-mediated isothermal amplification  real-time PCR

中文摘要:

      目的 建立食品过敏原牛奶成分LAMP(loop-mediated isothermal amplification)检测方法, 并与实时荧光PCR(real-time PCR)检测方法比对。方法 针对牛线粒体细胞色素b(cyt-b)基因设计LAMP引物并建立反应体系, 在特异性和灵敏度方面与real-time PCR检测方法比对。结果 本研究建立的LAMP方法检测9份不同品牌的牛奶和羊奶及其加工制品, 没有出现交叉反应, 具有良好的特异性。该方法的检测灵敏度为0.5%, 与real-time PCR方法检测灵敏度相当。检测了69份实际样品, 检测结果与real-time PCR检测结果一致。结论 本研究建立的食品过敏原牛奶成分LAMP检测方法简单经济, 检测结果可靠, 可有效缩短检测时间, 适用于过敏原牛奶成分的检测, 具有良好的应用前景。

英文摘要:

      Objective The loop-mediated isothermal amplification (LAMP) detection method was established for milk allergen testing and compared with the real-time PCR detection methods. Methods The cyt-b gene of bovine was used to design LAMP primers and then establish reaction system. The specificity and sensitivity of LAMP were compared with real-time PCR detection method. Results The specificity of LAMP method was tested by 9 different milk and goat milk products. The results showed that the LAMP method was highly specific to milk. No cross-reaction was founded. The detection limit of LAMP reached 0.5% in base-material addition test which was consistent with the real-time PCR method. Through 69 practical food samples testing, the LAMP results were consistent with real-time PCR results. Conclusion The LAMP detection method of milk allergen established in this study is simple, economical and reliable, which can effectively reduce the detection time and apply to the detection of milk allergen with good application prospects.