食品过敏原免疫传感检测技术研究

作为一个公共健康问题,食物过敏问题越来越引起人们的关注。目前,已确定的过敏性食物多达180种以上,联合国粮农组织报告了90%以上食物过敏原存在于牛奶、鸡蛋、鱼、甲壳类水产品、花生、大豆、坚果类及小麦8大类食物中。食品的过敏原是国内外检测、监控的一个重点、难点。根据检测目标物的不同,食物过敏原(Food allergen)检测技术主要有以下几种:分析全蛋白或酶解后肽段氨基酸序列的质谱技术、检测编码过敏原DNA片段的聚合酶链式反应(PCR)技术、分析过敏原性的免疫学检测技术。     

小麦过敏原脱敏技术研究进展

小麦是一种重要粮食,但小麦中含有一些过敏成份,其对人体的危害影响着小麦的开发与食用.综述小麦过敏原物质、过敏原检测技术、脱敏方法的研究进展,为小麦加工和使用提供指导.     

质谱技术在食物过敏原检测中的研究进展

食物过敏是当前食品安全领域比较突出的问题,相应的过敏原检测方法研究也越来越受重视。相比于传统的免疫学检测方法,质谱技术在检测加工后以及复杂基质中的食物过敏原中,具有高通量和高灵敏性等优点,被广泛应用于食物过敏原的检测。本文主要从定性和定量2方面介绍了质谱技术在食品过敏原检测中的研究进展。相关研究对牛乳、鸡蛋、小麦和榛子(坚果类)等主要食物过敏原均有涉及。在定性研究中,以肽质量指纹图谱法和肽碎片离子鉴定法为主,鉴定食物中的过敏原蛋白。在定量研究中,通过标记/无标记技术,也能够实现对微量的目标蛋白进行相对/绝对定量。质谱技术应用于食物过敏原检测中,将有助于提升过敏原检测能力,降低食物过敏安全风险。     

食物过敏原蛋白检测技术研究进展

目前食物过敏已经成为重要的食品安全问题,该文对食物中过敏原蛋白的检测方法进行综述,为不同食物中蛋白过敏原的检测提供依据。主要介绍了食物蛋白过敏原检测技术的新发展,包括酶联免疫吸附试验、免疫印迹法、免疫层析法、PCR法、SPR技术和质谱技术,并对食物过敏原检测方法的前景作出了展望。     

表面等离子共振技术及其在食物过敏原检测中的应用

表面等离子共振(SPR)技术是研究分子间及生物大分子稳定性的一种有效的非标记生物物理技术,并已经成功应用于使用简单、快速、无需标记的生物传感器。本文主要介绍了SPR的基本原理,SPR生物传感器的应用特点及设备以及在食物过敏原检测中的应用。已有的研究结果表明,SPR生物传感器在检测牛奶、坚果、鸡蛋和鱼制品中的主要过敏原等方面,具有灵敏度高、准确度高、检测快捷等特点,将发展成为食物过敏原检测的重要技术手段。     

食品过敏原澳洲坚果环介导等温扩增检测方法的建立与应用

根据澳洲坚果豌豆蛋白AMP2基因序列,利用设计软件Primer Explorer Version 4设计并筛选了食品过敏原澳洲坚果的环介导等温扩增引物,对反应体系和反应条件进行优化,建立澳洲坚果的环介导等温扩增检测方法,结果判断可采用实时荧光法和荧光染料终点显色法。对该方法进行了特异性、灵敏度、稳定性评价,结果显示：该方法能够特异性、灵敏、稳定地检测食品中的澳洲坚果成分,检测低限为0.5%。此外,对7 种市售食品样品的检测结果表明,该方法与食品标签标示的过敏原成分结果吻合率为100%,假阳性率和假阴性率均为0,在市售食品的过敏原成分检测上较商业化快速检测试纸条更加稳定可靠。     

物理法结合酶法脱除坚果类过敏原的技术研究

分别对4种坚果中的过敏蛋白进行分离提取,研究其蛋白特性,运用物理方法如高温、高压和微波超声波结合酶法作用于坚果过敏蛋白,探索脱除坚果类蛋白致敏性的最佳方法。结果表明:腰果、杏仁、核桃等坚果蛋白都使人产生过敏反应,经过高温、高压、超声波—微波联合萃取及蛋白酶处理后,均能使其致敏性明显降低。其中超声波—微波与酶法联合处理,超声波开启,作用温度90℃、时间240 s、微波功率700 W,0.25%胰蛋白酶处理5 min能完全脱除坚果类过敏原蛋白,是最佳脱敏方法。     

高静压在脱敏食品中的应用研究

高静压可以改变食物的结构和功能特性,掩盖食物过敏原的抗原表位,从而降低食物的过敏原性,被广泛应用于脱敏食品的加工过程中。阐述了食品脱敏的必要性,介绍了食品过敏原种类、作用机理及发病机理,综述了近几年高静压在食品脱敏中的应用。     

食品过敏原巴西坚果环介导等温扩增检测方法的建立与应用

根据巴西坚果2S白蛋白基因序列,利用设计软件Primer Explorer Version 4设计并筛选了食品过敏原巴西坚果的环介导等温扩增引物,对反应体系和反应条件进行了优化,建立了巴西坚果的环介导等温扩增检测方法,结果判断可采用实时荧光法和荧光染料终点显色法。以澳洲坚果、开心果、碧根果等17种常见坚果来验证方法的特异性;将巴西坚果DNA进行梯度稀释后验证方法的灵敏度;将0.5%、1%和1.5%3个浓度梯度的巴西坚果DNA重复检测20次来验证方法的稳定性。结果表明,本方法能够特异、灵敏、稳定地检测食品中的巴西坚果成分,检测低限为0.5%。此外,对7种市售食品样品的检测结果表明,该方法与食品标签标示的过敏原成分结果吻合率为100%,假阳性率和假阴性率均为0%。     

虾类过敏原及消减方法研究进展

虾及其制品味道鲜美,营养丰富,但却具有较高的致敏性。本文综述国内外有关虾类主要过敏原——原肌球蛋白的结构、表位预测与定位、变态反应原性检测及脱敏方法等方面的研究进展,为进一步研究虾类过敏原及其消减技术提供一定的参考。     

腰果过敏研究进展

腰果过敏是一种严重的树坚果过敏。近年来,随着腰果消费量迅速增长,腰果过敏发生率随之呈上升趋势,腰果正在成为一种重要的食物过敏原。少量的过敏原可导致严重的过敏反应,甚至危及性命。然而,目前腰果过敏显然是一个被严重低估的健康问题,尤其是对儿童。因此,社会迫切需要给予腰果过敏足够的重视,并有必要向公众广泛宣传腰果过敏的严重性。本文就腰果过敏的流行病学、过敏原成分、临床特点、诊断以及预防治疗等方面进行系统综述,旨在为腰果过敏的社会认识和深入研究提供科学信息。     

Processing effects on tree nut allergens: A review

“Tree nut” is a broad term for classification of nuts that include cashews, almonds, hazelnuts, etc. Reports of mild to adverse immune reactions following the consumption of these nuts has been on a rise in recent years. Currently, about 1.2–2% of the world's population suffer from sensitivity to tree nuts. The only solution is complete abstinence from the allergy causing tree nut which is not feasible in most cases due to issues like cross contamination or their presence in the form of hidden ingredients in processed foods. Various studies have shown that food processing can effectively vary the secondary structures of the allergenic protein which in turn influences their functional properties. But, the impact of these processing methods on tree nuts allergens is mixed. This review gives an update on the recent findings on how conventional and novel processing methods influence the tree nut allergens.     

One-step multiplex PCR method for the determination of pecan and Brazil nut allergens in food products

BACKGROUND: A one‐step polymerase chain reaction (PCR) method for the simultaneous detection of the major allergens of pecan and Brazil nuts was developed. Primer pairs for the amplification of partial sequences of genes encoding the allergens were designed and tested for their specificity on a range of food components. RESULTS: The targeted amplicon size was 173 bp of Ber e 1 gene of Brazil nuts and 72 bp of vicilin‐like seed storage protein gene in pecan nuts. The primer pair detecting the noncoding region of the chloroplast DNA was used as the internal control of amplification. The intrinsic detection limit of the PCR method was 100 pg mL^?1 pecan or Brazil nuts DNA. The practical detection limit was 0.1% w/w (1 g kg^?1). The method was applied for the investigation of 63 samples with the declaration of pecans, Brazil nuts, other different nut species or nuts generally. In 15 food samples pecans and Brazil nuts allergens were identified in the conformity with the food declaration. CONCLUSION: The presented multiplex PCR method is specific enough and can be used as a fast approach for the detection of major allergens of pecan or Brazil nuts in food. Copyright © 2011 Society of Chemical Industry     

Hazelnut Allergens: Molecular Characterization,Detection, and Clinical Relevance

In last few years, special attention has been given to food-induced allergies, in which hazelnut allergy is highlighted. Hazelnut is one of the most commonly consumed tree nuts, being largely used by the food industry in a variety of processed foods. It has been regarded as a food with potential health benefits, but also as a source of allergens capable of inducing mild to severe allergic reactions in sensitized individuals. Considering the great number of reports addressing hazelnut allergens, with an estimated increasing trend, this review intends to assemble all the relevant information available so far on the following main issues: prevalence of tree nut allergy, clinical threshold levels, molecular characterization of hazelnut allergens (Cor a 1, Cor a 2, Cor a 8, Cor a 9, Cor a 10, Cor a 11, Cor a 12, Cor a 14, and Cor a TLP) and their clinical relevance, and methodologies for detection of hazelnut allergens in foods. A comprehensive overview of the current data about the molecular characterization of hazelnut allergens is presented, relating to biochemical classification and biological function with clinical importance. Recent advances in hazelnut allergen detection methodologies are summarized and compared, including all the novel protein-based and DNA-based approaches.     

Pistachio nut allergy: An updated overview

Pistachio nut (Pistacia vera) is highly appreciated for its organoleptic characteristics and potential health benefits. However, this tree nut is also responsible for triggering moderate to severe IgE-mediated reactions in allergic individuals. Currently, pistachio nut allergy has gained some special attention, mainly due to its intrinsic relation with cashew nut allergy. Like for other nuts, the prevalence of pistachio nut allergy seems to be increasing at a global scale. Until now, there are five allergenic proteins officially listed for pistachio nut (Pis v 1, Pis v 2, Pis v 3, Pis v 4 and Pis v 5). Relevant data on their biochemical classification has become available, enabling establishing a correlation with the respective clinical symptoms. The establishment of an effective allergen risk assessment is a key issue for the food industry, policy makers and regulatory agencies. Thus, the availability of fast, specific and sensitive methods to detect trace amounts of allergens in processed foods is crucial. In the specific case of pistachio nut, there are some protein- and DNA-based methods for its detection/quantification in foods, which can aid to verify label information. Accordingly, all relevant research advances on this topic were summarised, updated and critically discussed in this review.     

甲壳类水产食物致敏原及其分子结构的研究进展

甲壳类水产品是亚洲地区最主要的致敏食物。食物过敏主要由摄入的食品致敏原引起,而食物致敏原的结构不仅与抗原表位的形成有关,也决定了其在食品加工和体内消化过程的稳定性。本文针对甲壳类水产品,首先阐述了甲壳类水产食物过敏及致敏原的研究进展;进一步对甲壳类水产食物致敏原的分子结构及其与致敏性的构效关系展开论述,并基于此提出了致敏原脱敏策略。目前已鉴定的甲壳类水产食物致敏原大部分属于肌动蛋白结合蛋白、磷酸原激酶和EF手型钙离子结合蛋白3个家族。针对不同蛋白质家族致敏原的结构特点,通过食品加工技术消减致敏性或利用分子生物学手段获得低致敏性衍生物都是防控食物过敏的重要途径。但甲壳类水产食物致敏原的结构信息有待完善,其与致敏性的构效关系也仍是未来致敏原研究的重要方向。本文综合阐述了甲壳类水产食物致敏原结构特性及其与致敏性的构效关系,以期为甲壳类水产食物致敏原的系统研究和食品脱敏方法的建立提供思路。     

A Rapid Size-Exclusion Solid-Phase Extraction Step for Enhanced Sensitivity in Multi-Allergen Determination in Dark Chocolate and Biscuits by Liquid Chromatography–Tandem Mass Spectrometry

Enhanced sensitivity for the simultaneous determination of five nut allergens in biscuit and in dark chocolate complex matrices was obtained by introduction of a rapid size-exclusion solid-phase extraction-based step before liquid chromatography–electrospray ionization-tandem mass spectrometry (LC-ESI-MS²) analysis. A very fast and efficient separation (<12 min) of marker peptides with selected reaction monitoring detection was obtained. Limits of detection in the 0.1–1.3 mg nut/kg and 5–15 mg nut/kg ranges for biscuit and dark chocolate samples as well as high recoveries (84(±6)–106(±4)% for biscuits and 98(±5)–108(±6)% for dark chocolate) proved the excellent capabilities of the exploited sample treatment method combined with the LC-MS² analysis. Good precision in terms of intra- and inter-day repeatability was calculated, being always lower than 19 % (n?=?75). Linearity was demonstrated up to four and three orders of magnitude for biscuit and dark chocolate, respectively. Finally, the validated method was successfully applied to the investigation of hidden nut trace allergens in commercially available biscuits and chocolates of different brands aiming to ascertain possible discrepancies between allergen content and food allergen labelling.     

Application of real-time PCR for tree nut allergen detection in processed foods

Abstract

Currently, food allergies are an important health concern worldwide. The presence of undeclared allergenic ingredients or the presence of traces of allergens due to accidental contamination during food processing poses a great health risk to sensitized individuals. Therefore, reliable analytical methods are required to detect and identify allergenic ingredients in food products. Real-time PCR allowed a specific and accurate amplification of allergen sequences. Some processing methods could induce the fragmentation and/or degradation of genomic DNA and some studies have been performed to analyze the effect of processing on the detection of different targets, as thermal treatment, with and without applying pressure. In this review, we give an updated overview of the applications of real-time PCR for the detection of allergens of tree nut in processed food products. The different variables that contribute to the performance of PCR methodology for allergen detection are also review and discussed.     

食品中过敏原及其检测方法的研究进展

近年来食物过敏作为食品安全的热点问题在全球引起了广泛关注。目前食物过敏仍无有效的根治方法,严格避免摄入致敏食品仍是过敏性疾病防治的最有效方式。因此,食品中过敏原的识别鉴定、性质分析及检测方法的研究至关重要。本文针对八大类过敏食物,从分子结构、免疫学特性等方面介绍了常见食物过敏原的研究进展;归纳了现阶段用于食品中过敏原分析的常规检测方法,以及在此基础上发展而来的新兴检测丰富,并比较分析了不同检测技术方法的优缺点。相比于植物源性过敏原,动物源性过敏原的相关研究有待深入,尤其是蛋白家族的确定及空间结构解析等是未来动物源性过敏原研究的重要方向;目前食品中常见过敏原的常规检测方法及新兴检测方法依然存在检测效率和准确性不高、成本昂贵等局限性,建立高效、准确、低成本的过敏原检测方法仍是该研究领域发展的重要趋势。     

坚果品质检测方法研究进展

坚果含有优质蛋白质、脂肪等丰富的营养物质,深受消费者喜爱。由于富含不饱和脂肪酸及油脂,坚果的品质易受贮藏条件、加工方式及包装方式等外部因素的影响而发生变化,不当的处理会加速其氧化哈败,甚至产生对人体有害的毒素。另一方面,坚果品质受产地、品种、土壤环境及遗传因素等方面的影响表现出不同的外观性状及理化特征。随着科学技术的发展,坚果品质的检测趋向于无损、快速,电子鼻在这方面有着突出的优势,因此电子鼻对坚果品质的无损检测成为研究的新热点。本文从口感、色泽、风味、外观性状等感官检测,营养品质（蛋白质、脂肪等）、油脂品质（酸价、过氧化值等）等理化检测,光谱分析技术（近红外光谱、高光谱等）、电子鼻技术等快速检测方法综述了坚果品质检测方法的研究现状,为坚果品质的监控和检测提供技术参考。