基于多重连接依赖探针扩增(MLPA)技术检测加工食品中过敏原成分  

Detection of the Allergens in Processed Foods Based on Multiple Ligation-dependent Probe Amplification(MLPA)

作　　者：刘艳 王鸣秋 李诗瑶 张涛 何名扬 朱必婷 林津 徐芬 张莉 LIU Yan;WANG Mingqiu;LI Shiyao;ZHANG Tao;HE Mingyang;ZHU Biting;LIN Jin;XU Fen;ZHANG Li(Hubei Provincial Institute for Food Supervision and Test,Key Laboratory of Detection Technology of Focus Chemical Hazards in Animal-derived Food for State Market Regulation,Hubei Provincial Engineering and Technology Research Center for Food Quality and Safety Test,Wuhan 430075,China)

机构地区：[1]湖北省食品质量安全监督检验研究院,国家市场监管重点实验室(动物源性食品中重点化学危害物检测技术),湖北省食品质量安全检测工程技术研究中心,湖北武汉430075

出　　处：《现代食品科技》2023年第6期290-297,共8页Modern Food Science and Technology

基　　金：湖北省重点研发计划项目(2020BCA091);国家市场监督管理总局科技计划项目(2020MK067)。

摘　　要：该研究基于多重连接探针扩增技术(Multiplex Ligation-dependent Probe Amplification,MLPA)同时检测开心果、巴西坚果、芹菜、麸质、夏威夷果、芝麻、榛子、大豆、花生、葵花籽、核桃、腰果、杏仁、芥末等14种植物源性的过敏原成分,针对ITS序列设计特异性杂交探针,样本核酸经95℃变性后,与探针进行特异性结合,经连接和PCR扩增反应得到不同大小的目标片段,通过毛细管电泳分析目标片段的有无来判断是否含有待测过敏原成分。利用混合探针体系检测单一模板只能扩增出单一扩增峰,表明探针具有高特异性,检测限结果表明,MLPA扩增最低可检出的DNA质量浓度为1ng/μL。通过20份实际样本的检测,证明该研究基于MLPA建立的加工食品中过敏原成分的检测方法,具有特异性强,灵敏度高的特点,可以应用于食品安全监管工作。In this study,multiplex ligation-dependent probe amplification(MLPA)was applied to simultaneously detect 14 plant-derived allergens from pistachio,Brazil nut,celery,gluten,macadamia nut,sesame,hazelnut,soybean,peanut,sunflower seed,walnut,cashew,almond and mustard.Specific hybridization probes were designed for ITS sequences.After the sample nucleic acid was denatured at 95,specific℃binding was carried out with the probes,and target fragments of different sizes were obtained through ligation and PCR amplification reaction.The target fragments were analyzed by capillary electrophoresis to determine whether or not the allergens were present.Using the mixed probe system to detect a single template canonly amplifya single amplification peak,indicating that the probe had high specificity.The detection limit results showed that the lowest detectable DNA concentration of MLPA amplification was 1 ng/μL.Through the detection of 20 actual samples,it is proven that the detection method of allergens in processed foods established in this study on the basis of MLPA has the characteristics of high specificity and high sensitivity,and can be applied to food safety supervision.

关 键 词：多重连接探针扩增技术 过敏原 特异性 灵敏度 

分 类 号：TS207.3[轻工技术与工程—食品科学]