实时LAMP法快速检测食用植物油中的转基因成分CaMV-35S

环介导等温扩增技术(Loop-mediated isothermal amplification,LAMP)是一项新的DNA扩增技术,近年来已被成功应用于转基因作物中外源基因的检测分析。本文针对花椰菜花叶病毒35 S启动子(CaMV-35S)设计引物以及有效提取目标DNA后,首次建立了食用植物油中CaMV-35S启动子的LAMP检测方法。通过在DNA提取过程中加入正己烷乳化和加入共沉淀剂,获得了可以进行LAMP扩增的DNA片段,采用LAMP实时浊度法和染色法对扩增产物进行比较分析。着重对FIP/BIP引物、甜菜碱和Mg2+浓度等反应参数进行了优化。结果表明,LAMP方法能够在56 min内特异性地检测到CaMV-35S启动子,检测灵敏度比常规PCR高10倍,而且不需要特别的仪器设备,扩增产物可通过观察实时浊度曲线或通过SYBR Green I染色后借助肉眼对检测结果进行判断。该方法快速高效、操作简便、灵敏度高、检测结果准确,适合食用植物油中转基因成分的快速检测。     

Effect of non‐isothermal processing and moisture content on the anthocyanin degradation and colour kinetics of cherry pomace

Anthocyanins (ACY) and colour changes in cherry pomace under non‐isothermal processing were investigated. Pomace at moisture levels of 70% (MC‐70), 41% (MC‐41) and 25% (MC‐25) was heated at 126.7 °C in a retort for 25, 40 and 60 min. Total ACY, Hunter colour values, total colour difference (ΔE), chroma, hue angle (h°) and browning index (BI) were analysed. Thermal degradation kinetics for colour parameters were determined using zero‐ and first‐order models. ACY degradation increased with heating time and ranged from 34 to 68% for 25 and 60 min heating, respectively. The half‐life of ACY was 38, 33 and 27 min for MC‐70, MC‐41 and MC‐25 pomace, respectively. The ΔE increased with increasing heating time, whereas BI exhibited an inverse trend. Except for ?E for MC‐70, the zero‐order kinetic model showed better fit (R² = 0.85–0.97) to experimental data than the first‐order kinetic model for Hunter colour b values and ?E.     

适配体功能化的DNA水凝胶的制备及其在食品安全小分子快速检测中的应用

DNA水凝胶是具有三维聚合物网络的高保水性材料。研究人员设计了多种DNA水凝胶交联制备方法,并通过向其中引入其他功能分子或与其他类型的功能材料相互结合,构建了具有优异性能的DNA水凝胶,受到了广泛关注。适配体是基于指数富集的配体系统进化(systematicevolutionofligandsbyexponential enrichment,SELEX)技术从随机寡核苷酸文库中筛选获得的对目标物质具有良好特异性与亲和力的寡核苷酸序列。适配体功能化的DNA水凝胶具有靶向范围广、稳定性好、易于修饰、操作简单和成本低等优点,得到了广泛应用。本文概述了构建适配体功能化的DNA水凝胶的基本设计原则与分类,重点介绍了适配体功能化的DNA水凝胶在食品安全检测中的最新策略,最后,讨论了适配体功能化的DNA水凝胶面临的挑战以及对未来的展望,旨在为其在食品安全领域的应用提供参考。     

基于可视化环介导等温扩增技术快速检测金黄色葡萄球菌

应用环介导等温扩增技术（loop-mediated isothermal amplification,LAMP）建立基于颜色判定的快速、灵敏的金黄色葡萄球菌检测方法。根据金黄色葡萄球菌特异性靶基因SAR0395设计LAMP引物,建立可视化LAMP检测方法。优化的反应体系为：1.6 μmol/L内引物（FIP和BIP）,0.2 μmol/L外引物（F3和B3）,4.0 mmol/L MgSO4,1.0 mmol/L dNTPs,0.4 U/μL Bst 2.0 WarmStart? DNA聚合酶,120 μmol/L羟基萘酚蓝,扩增温度60 ℃,扩增时间60 min。在可视化LAMP体系中添加两条环引物（LF和LB）反应时间可缩短至20 min。该可视化LAMP反应60 min,其基因组灵敏度可以达到0.010 3 fg/μL,菌落灵敏度可以达到1.9 CFU/mL;利用46 株金黄色葡萄球菌和24 株非金黄色葡萄球菌证实该可视化LAMP具有良好的特异性和可靠性。本研究中建立的可视化LAMP可为金黄色葡萄球菌的快速检测提供一种新的技术。     

重组酶聚合酶恒温扩增结合乳胶微球试纸条快速检测金黄色葡萄球菌

通过建立一种将重组酶聚合酶恒温扩增(recombinase polymerase amplification,RPA)与乳胶微球试纸条(latex microsphere test strips,LMTS)相结合的方法,快速检测金黄色葡萄球菌。根据金黄色葡萄球菌保守区序列(nuc)设计特异性引物,经RPA扩增后用LMTS检测,确定RPA-LMTS检测金黄色葡萄球菌的灵敏度和特异性。灵敏度结果显示RPA-LMTS检测限为500 fg DNA和1.2×101 CFU/mL纯菌液;特异性结果表明与沙门氏菌、大肠杆菌O157∶H7、志贺氏菌、产气肠杆菌和单增李斯特菌无交叉反应,特异性良好;用食物样品评估RPA-LMTS检测效果,结果显示经过增菌3小时后,RPA-LMTS可检测1.2×100 CFU/mL(g)的金黄色葡萄球菌。     

等温扩增技术在食源性致病菌检测中的应用进展

等温扩增技术是近年来迅速发展的一类核酸扩增技术。相比于PCR,该技术具有高特异性、高敏感性、简单、便捷及低成本的特点。目前,核酸等温扩增技术在感染性疾病的诊断、基因多态性分析、疫情防治等方面已经有广泛应用,也有文献报道了其在细菌、病毒等病原体检测方面的应用。食源性致病菌和环境中的病原体检测等领域中,等温扩增技术展现了广阔的应用潜力,有望发展成为食源性致病菌检测的重要方法。本文综合国内外文献报道,对环介导等温扩增、依赖解旋酶等温扩增、依赖核酸序列等温扩增、切刻内切酶核酸恒温扩增、交叉引物等温扩增、链置换等温扩增、SmartAmp技术等一系列等温扩增技术的原理、特性及其在食源性致病菌检测中的应用情况做一综述,从而为该技术在食源性致病菌检测的实际应用中提供参考和依据。     

Some interrelated thermophysical properties of liquid water and ice. I. A user-friendly modeling review for food high-pressure processing

A bibliographic search yielded a set of empirical equations that constitute an easy method for the calculation of some thermophysical properties of both liquid water and ice I, properties that are involved in the modeling of thermal processes in the high-pressure domain, as required in the design of new high-pressure food processes. These properties, closely interrelated in their physical derivation and experimental measurement, are specific volume, specific isobaric heat capacity, thermal expansion coefficient, and isothermal compressibility coefficient. Where no single equation was found, an alternative method for calculation is proposed. Keeping in mind the intended applications and considering the availability of both experimental data and empirical equations, the limits for the set of equations where set in -40 to 120 degrees C and 0 to 500 MPa for liquid water and -30 to 0 degrees C and 0 to 210 MPa for ice I. The equations and methods selected for each property are described and their results analyzed. Their good agreement with many existing experimental data is discussed. In addition, the routines implemented for the calculation of these properties after the described equations are made available in the public domain.     

Current and emerging molecular diagnostic technologies applicable to bacterial food safety

There is an increasing need for rapid test methods to certify the quality and safety of food products. Current tests applied for the microbiological assessment of food products are based on standard approved culture-based isolation methods and can take several days to yield results. Nucleic acid diagnostic (NAD) tests for the identification of bacterial foodborne pathogens employing in vitro amplification technologies are capable of sensitive and specific detection of single or multiple pathogens in foods in a shorter timeframe than traditional methods. New developments in molecular biosensors have the potential to provide at-line bioanalysis, whereas microarray-based technologies may in the future be the NAD platforms of choice for multiple pathogen detection and identification. This article reviews current and emerging NAD platforms for foodborne bacterial pathogens that have the potential to impact food safety.     

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。