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

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。

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 spe-cificity 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, economi-cal and reliable, which can effectively reduce the detection time and apply to the detection of milk aller-gen with good application prospects.