食品中芥末过敏原成分LAMP检测方法的建立  

Establishment of a Loop-Mediated Isothermal Amplification Method for Detection of Mustard Allergen in Foods

作　　者：程晋霞[1,2] 周熙诚 马丹[1] 刘莉[1] 曾静[1] 

机构地区：[1]北京出入境检验检疫局,北京100026 [2]北京农学院,北京102206

出　　处：《食品科学》2014年第20期148-152,共5页Food Science

基　　金："十二五"国家科技支撑计划项目(2011BAK10B03)

摘　　要：目的:建立食品过敏原芥末成分环介导等温扩增(loop-mediated isothermal amplification,LAMP)检测方法,并与实时荧光-聚合酶链式反应(real-time polymerase chain reaction,RT-PCR)检测方法比对。方法:针对白芥的主要过敏原基因Sin A1,设计LAMP引物并建立反应体系,在特异性和灵敏度方面与RT-PCR检测方法比对。结果:建立的LAMP检测方法,经特异性验证,与所测试的13种植物,无交叉反应。通过添加实验,方法的检测灵敏度为0.5%,与RT-PCR方法检测灵敏度相当。检测了25份实际样品,检测结果与RT-PCR检测结果一致。结论:建立的食品过敏原芥末成分LAMP检测方法简单经济,检测结果可靠,可有效缩短检测时间,适用于芥末过敏原成分的检测,具有良好的应用前景。Objective: A loop-mediated isothermal amplification(LAMP) method was established for the detection of mustard allergen in foods and was compared with real-time polymerase chain reaction(RT-PCR). Methods: The Sin A1 gene of Sinapis alba was used to design LAMP primers. The specificity and sensitivity of LAMP were compared with those of RT-PCR. Results: The specificity of LAMP method was tested with 13 different crops. The results showed that the LAMP method was highly specific to mustard. No cross-reaction was found. The limit of detection(LOD) for LAMP was 0.5%in base-material addition test, which was consistent with that of the RT-PCR method. For 25 real food samples, the LAMP results were consistent with the RT-PCR results. Conclusion: The LAMP method of established in this study was simple, economical and reliable and could effectively reduce the detection time.

关 键 词：芥末 过敏原 环介导等温扩增 实时荧光-聚合酶链式反应 

分 类 号：O657.3[理学—分析化学] TS207.3[理学—化学]