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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 To establish a loop-mediated isothermal amplification (LAMP) method for food allergen and compare it with real-time PCR. Methods The LAMP primer was designed according to the cytochrome b gene of bovine mitochondria and the reaction system was established. The specificity and sensitivity were compared with the real-time PCR assay. Results The LAMP method established in this study detected 9 different brands of milk and goat milk and their processed products without any cross-reaction and had good specificity. The detection sensitivity of this method is 0.5%, which is equivalent to the sensitivity of real-time PCR method. 69 real samples were tested and the results were in good agreement with real-time PCR. Conclusion The milk allergen LAMP assay established in this study is simple and economical, reliable test results, which can effectively shorten the test time and is suitable for the detection of allergen milk components, which has a good application prospect.