Objective: A multiplex fluorescence PCR method based on TaqMan probe was developed for simultaneous detection of soybean and wheat allergens in food. Methods: TaqMan probe dual PCR was designed and established based on wheat gliadin gene and soybean lectin gene, respectively. The soybean Lectin gene was detected with FAM marker, and the wheat gliadin gene was detected with HEX marker. Moreover, the eukaryotic 18S rRNA was used as internal reference gene to ensure the effectiveness of the detection system. Results: The multiplex fluorescence PCR method for detection of soybean and wheat allergens created in this study had no cross-reaction to ingredients other organisms showing strong specificity. The detection limit of soybean and wheat mixture was up to 0.01%(mass ratio), and the detection sensitivity was high. Conclusion: The established TaqMan probe real-time multiplex detection system for soybean and wheat allergens has the characteristics of good specificity and high sensitivity, which can be used for simultaneous and rapid detection of soybean and wheat allergens in food.

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[1] 黄霆钧,张欣琪,刘安游,等.我国食品过敏原标准体系构建研究[J].中国标准化,2020(3):129-132,164.HUANG T J,ZHANG X Q,LIU A Y,et al.Research on the construction of food allergen standards system in china[J].China Standardization,2020(3):129-132,164.
[2] 宁晖,房方.食品过敏法规及其检测技术现状[J].食品安全质量检测学报,2020,11(12):3 768-3 772.NING H,FANG F.Regulations and current detection technology of food allergen[J].Journal of Food Safety & Quality,2020,11(12):3 768-3 772.
[3] 王雅清,倪皓洁,李华韬,等.食物过敏原检测技术研究进展[J].食品工业科技,2019,40(13):264-270.WANG Y Q,NI H J,LI H T,et al.Research advance in detection technology for food allergen[J].Science and Technology of Food Industry,2019,40(13):264-270.
[4] 张晶宇.基于蛋白质组学的鱼类过敏原检测方法研究[D].大连:大连海洋大学,2019:7-22.ZHANG J Y.Proteomics-based detection of fish allergens[D].Dalian:Dalian Ocean University,2019:7-22.
[5] 郑义成,华萍,杨安树,等.食物中过敏原检测技术研究进展[J].食品科学,2010,31(21):417-420.ZHENG Y C,HUA P,YANG A S,et al.Research advance in detection technologies for allergen in food[J].Food Science,2010,31(21):417-420.
[6] 郭颖慧,霍胜楠,孟静,等.食品过敏原检测技术研究进展[J].食品安全质量检测学报,2019,31(21):417-420.GUO Y H,HUO S N,MENG J,et al.Research progress of detection technologies for allergen in food[J].Journal of Food Safety & Quality,2019,31(21):417-420.
[7] WALCZYK N E,SMITH PM C,TOVEY E R,et al.Peanut protein extraction conditions strongly influence yield of allergens Ara h 1 and 2 and sensitivity of immunoassays[J].Food Chemistry,2017,221:335-344.
[8] LUIS A,GONZALEZ I,GARCIA T,et al.Determination of food authenticity by enzyme-linked immunosorbent assay(ELISA)[J].Food Control,2008,19(1):l-8.
[9] BENREJEB S,ABBOTT M,DAVIES D,et al.Multi-allergen screening immunoassay for the detection of protein markers of peanut and four tree nuts in chocolate[J].Food Addit Contam,2005,22(8):709-715.
[10] POLLET J,DELPORT F,JANSSEN K P F,et al.Fast and accurate peanut allergen detection with nanobead enhanced optical fiber SPR biosensor[J].Talanta,2011,83(5):1 436-1 441.
[11] JOHNSON P E,RIGBY N M,DAINTY J R,et al.A multi-laboratory evaluation of a clinically-validated incurred quality control material for analysis of allergens in food[J].Food Chem,2014,148:30-36.
[12] CUCU T,DE MEULENAER B,KERKAERT B,et al.MALDI based identification of whey protein derived tryptic marker peptides that resist protein glycation[J].Food Res Inter,2012,47(1):23-30.
[13] MONACI L,VISCONTI A.immunochemical and DNA-based methods in food allergen analysis and quality assurance perspectives[J].Trends in Food Science and Technology,2010,21(6):272-283.
[14] GARCIA-RUIZ C,TORRE M,MARINA M L.Analysis of bovine whey proteins in soybean dairy-like products by capillary electrophoresis[J].J Chromatogr A,2009,859(1):77-86.
[15] 徐双双,王尉,陈尔凝,等.电泳法在食品过敏原检测中的应用[J].食品安全质量检测学报,2019,10(11):3 436-3 443.XU S S,WANG W,CHEN E N,et al.Application of electrophoresis in the detection of food allergens[J].Journal of Food Safety & Quality,2019,10(11):3 436-3 443.
[16] SIVAGURU M,BALUSKA F,VOLKMANN D,et al.Impacts of aluminum on the cytoskeleton of the maize root apex.short-term effects on the distal part of the transition zone[J].Plant Physiology,2009,119:1 073-1 082.
[17] SAMAC D A,TESFAYE M.Plant improvement for tolerance to aluminum in acid soils:A review[J].Plant Cell Tissue & Organ Culture,2003,75:189-207.
[18] SAMUELS T D,KUCUKAKYUZ K,RINCON-ZACHARY M.Al partitioning patterns and root growth as related to Al sensitivity and Al tolerance in wheat[J].Plant physiology,2007,113:527-534.
[19] 陈庆山,刘春燕,刘迎雪,等.核酸体外扩增技术[J].中国生物工程杂志,2004,24(5):10-14.CHEN Q S,LIU C Y,LIU Y X,et al.Progress of nucleic acid amplification technologies[J].China Biotechnology,2004,24(5):10-14.
[20] 董旭婉.食品中植物源性成分数字PCR定量检测方法研究[D].广州:华南理工大学,2020:29-59.DUNG X W.Study on quantitative detection of plant-derived components in food by digital PCR[D].Guangzhou:South China University of Technology,2020:29-59.
[21] 兰海鸥.大豆、核桃、花生过敏原基因三重实时荧光检测方法的建立及应用[D].兰州:西北民族大学,2020:22-43.LAN H O.Establishment and application of triple real-time fluorescence detection method for allergen genes in soybean,walnut and peanut[D].Lanzhou:Northwest University for nationalitie,2020:22-43.
[22] ZAHRADNIK C,MARTZY R,MACH R L,et al.Detection of the food allergen celery via loop-mediated isothermal amplification technique[J].Anal Bio Chem,2014,406(27):6 827-6 833.
[23] 陈云芳,詹国辉,高渊.核酸恒温扩增技术的原理及其应用[J].西南林学院学报,2010,30(S1):30-32,40.CHEN Y F,ZHAN G H,GAO Y.Principle and application of isothermal DNA amplification[J].Journal of Southwest Forestry College,2010,30(S1):30-32,40.
[24] KOLLMEIER M,FELLE H H,HORST W J.Genotypical differences in aluminum resistance of maize are expressed in the distal part of the transition zone.Is reduced basipetal auxin flow involved in inhibition of root elongation by aluminum[J].Plant Physiolog,2000,122:945-956.
[25] BLARNEY F P C,NISHIZAWA N K,YOSHIMURA E.Timing,magnitude,and location of initial soluble aluminum injuries to mungbean roots[J].Soil Science & Plant Nutritio,2004,50:67-76.
[26] ZHENG S J,YANG J L.Target sites of aluminum phytotoxicity[J].Biologia Plantaru,.2005,49:321-331.
[27] 任易婕,霍胜楠,翟清燕,等.膜芯片技术对牛、羊、牦牛、驴肉源食品的掺伪鉴别[J].肉类研究,2019,33(6):33-38.REN Y J,HUO S N,ZHAI Q Y,et al.Identification of adulterated meat from cattle,sheep,yak and donkey by membrane chip technology[J].Meat Research,2019,33(6):33-38.

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