质谱法检测食源性致病菌技术研究进展

食源性致病菌存在广泛,能够引起人类的疾病甚至死亡,研究发现超过一半的食品安全问题来源于食源性致病菌的污染。如何快速有效地检测出食源性致病菌是预防和控制食品安全问题的关键环节。系统地介绍了检测食源性致病菌的方法,包括传统培养法、代谢学法、分子生物学法、免疫学方法等传统方法以及新兴的质谱法。质谱法有检测效率高、操作简便、灵敏度高等优点,着重对质谱法的原理、应用以及未来的发展趋势进行了阐述,以期为该技术的研究开发和推广应用提供参考。     

显色培养基在几种食源性致病菌快速检测中的应用

用显色培养基鉴定微生物是一种新的微生物快速检测技术,该技术以生化反应为基础,通过在培养基中加入细菌特异性酶的显色底物直接根据菌落颜色对菌种作出鉴定。常见食源性致病菌检测中,李斯特菌显色培养基（BCM^TM Listeria mormcytogenes,Rapid’LMONO agar．CHROMagar^TM Listeria）、大肠杆菌显色培养基（CHROMagar^TM Ecoli）、沙门氏菌显色培养基（Rambach agar）、金黄色葡萄球菌显色培养基（CHROMagar Staphylococcus aureus）等已被广泛应用于食品、医药和环境监测等领域,极大提高了微生物检测的效率。但是,显色培养基也存在一些假阳（阴）性等问题,其设计尚待优化。     

PCR技术检测食源性致病菌的研究进展

食源性致病菌的检测技术是食源性疾病预防与控制的关键环节。PCR是近年来广泛应用于食源性致病菌快速检测的方法之一。在食源性致病菌中,用于PCR检测的靶基因包括各种毒力基因、酶基因及特异性鉴别基因。这些靶基因的发现推动了食源性致病菌PCR快速检测的发展。     

食源性致病菌检测免疫传感器研究进展

食源性致病菌是造成食品安全事件的主要原因之一,因此其检测方法已成为人们研究的热点.食源性致病菌的检测方法主要有病原体培养法、免疫学方法、核酸检测和生物传感器等.其中,免疫传感器基于抗原抗体特异性结合,整合光学、电化学等多学科交叉技术,具有特异性强、检测速度快等特点.本文对比食源性致病菌传统检测方法,综述了近年来免疫传感...     

食源性致病菌检测技术的发展及研究现状

食源性疾病对人类健康产生越来越大的威胁,且因为抗菌类药物的广泛使用,食源性致病菌出现多重耐药现象。作为预防与控制食源性致病菌的关键环节,食源性致病菌快速检测技术的开发尤为重要。传统检测技术包括微生物培养法、免疫学检测技术和分子检测技术,存在周期长、对检测人员和检测环境要求高等不足,如何将不同检测技术的优点集于一体而规避相应的缺点是突破方向。随着生物科技的发展,新型检测技术逐渐兴起,如基于光学、电化学的生物传感技术或多种技术结合的新应用等。本文综述了常见食源性致病菌的生理特性及相应感染疾病,讨论了传统检测方法优缺点,并详细介绍了新型生物传感器检测技术的发展及现状,以期为开发更加便捷、准确、灵敏的食源性致病菌现场检测技术提供参考。     

食源性致病菌多重分子生物学检测技术研究进展

快速、可靠的食源性致病菌高通量检测方法对于确保食品安全具有重要意义,近年基于DNA水平的多重分子生物学检测技术迅速发展,针对各种不同的食源性致病菌建立了多种多重分子检测技术,包括多重PCR、多重实时荧光PCR以及基因芯片等。对这些多重分子检测技术的最新研究进展作一综述,并且建议在今后该技术的研究中,仍需要在食品中多种致病菌同时选择性增菌培养、亚致死损伤修复以及检测内标的构建等方面取得突破,从而能够更好地实现食源性致病菌的高通量检测。     

生物传感器在食源性致病菌检测中应用的研究进展

食源性致病菌作为引起食源性疾病的主要因素,受到人们的高度重视,发展简便、快速、高灵敏度和低成本的食源性致病菌检测方法对降低食源性疾病发病率具有重要意义。生物传感器技术是一种由多学科交叉渗透发展形成的全新微量分析技术,具有灵敏度高、分析速度快等特点,被广泛应用于食源性致病菌的检测。文中介绍了生物传感器的基本原理,综述了常见的生物传感器在食源性致病菌检测中的应用,并对其发展趋势进行了展望。     

显色培养基在几种食源性致病菌快速检测中的应用*

用显色培养基鉴定微生物是一种新的微生物快速检测技术,该技术以生化反应为基础,通过在培养基中加入细菌特异性酶的显色底物直接根据菌落颜色对菌种作出鉴定。常见食源性致病菌检测中,李斯特菌显色培养基(BCM^TMListeriamonocytogenes,Rapid’LMONOagar,CHROMagar^TMListeria)、大肠杆菌显色培养基(CHROMagarTMEcoli)、沙门氏菌显色培养基(Rambachagar)、金黄色葡萄球菌显     

食源性致病菌多重PCR快速检测方法建立与应用

利用PCR技术,建立多组多重食源性致病菌PCR快速检测方法。设计受试菌特异性引物,反应体系中加入多对引物和多种DNA模板,采用正交试验优化PCR反应条件,进行特异性引物的PCR扩增。建立了多组多重食源性致病菌PCR快速检测方法,方法中所检测受试菌株和模拟样品均出现特异性扩增条带,结果与实际相符。所建立多组多重PCR快速检测体系符合设计要求,可以应用于食源性突发公共卫生事件的应急检测和日常样品检测工作。     

食源性致病菌核酸标准物质的研究进展

核酸检测试剂盒在微生物快速检测领域飞速发展,使检测过程具有操作简单、成本低、便于现场检测的优势,食源性致病菌核酸检测试剂盒在制备过程中需要通过计量溯源以确保其定性及定量的测量能力。核酸标准物质（nucleic acid reference materials,NARMs）应用于临床卫生、食品、仪器检定/校准以及核酸检测等多个领域,通过片段化分割遗传物质,最大程度地保障生物安全性,满足质量评价和计量溯源的需求,能够对核酸检测试剂盒进行有效质控,为核酸检测试剂盒的量值评价提供精准的“标尺”。综述了我国NARMs的研制方法,重点阐述了目前食源性致病菌的研究现状以及在国家标准物质资源共享平台的应用情况,分析了面临的技术挑战以及未来核酸计量工作发展的可能性,以期为核酸检测行业的早期诊断和防控提供参考。     

基于CRISPR/Cas系统的纸基分析在快速检测食源性致病菌中的应用

由食源性致病菌引起的食品安全事件严重影响人类健康,开发针对食源性致病菌的快速检测技术十分必要。成簇间隔短回文重复序列（clustered regularly interspaced short palindromic repeats,CRISPR）及相关蛋白（CRISPR-associated protein,Cas）是原核生物的适应性免疫系统,具有特异性识别并切割核酸序列的功能。纸基分析方法作为一种简便性好、成本低廉的分析检测工具,在快速检测领域展现出良好的前景。因此,将CRISPR/Cas系统的高效识别能力和纸基分析方法的简便性相结合可实现对食源性致病菌的快速灵敏检测。本文简要介绍了CRISPR/Cas系统用于核酸检测的概况,对第二类单Cas效应蛋白系统的特点及原理进行概述,重点综述基于CRISPR/Cas系统的试纸分析、侧向流动分析和纸基微流控装置在检测食源性致病菌方面的应用,并讨论了CRISPR/Cas系统结合纸基分析建立检测方法的优势、当前的挑战及未来的发展前景。     

The application and research progress of bacteriophages in food safety

The abuse of antibiotics and the emergence of drug-resistant bacteria aggravate the problem of food safety. Finding safe and efficient antibiotic substitutes is an inevitable demand for ensuring the safety of animal-derived food. Bacteriophages are a kind of virus that can infect bacteria, fungi or actinomycetes. They have advantages of simple structure, strong specificity and nontoxic side effects for the human body. Bacteriophages can not only differentiate live cells from dead ones but also detect bacteria in a viable but nonculturable state. These characteristics make bacteriophages more and more widely used in the food industry. This paper describes the concept and characteristics of bacteriophages, and introduces the application of bacteriophages in preharvest production, food processing, storage and sales. Several methods of using bacteriophages to detect foodborne pathogens are listed. Finally, the advantages and limitations of bacteriophages in the food industry are summarized, and the application prospect of bacteriophages in the food industry is discussed.     

合成生物学细胞传感技术在食品安全快速检测中的应用

合成生物学细胞传感技术为快速、现场检测食品污染物提供了一种新型替代方法。由于细胞内环境相对稳定,合成生物学细胞传感器有较强的抗干扰能力;由于细胞能够通过自我复制而实现增殖,细胞传感器在生产上具有简单、廉价、快速的特点,因此在食品安全快速检测中具有良好的应用前景。本文综述了合成生物学细胞传感器核心元件的组成、构建方法和类型,介绍了多功能细胞传感器的合成生物学基因回路,列举了细胞传感器在食品安全快速检测中的商业化应用前景,并阐述了细胞传感器在食品安全快速检测中的挑战和发展趋势。     

Reduction of Salmonella on alfalfa seeds using peroxyacetic acid and a commercial seed washer is as effective as treatment with 20 000 ppm of Ca(OCl)2

Aims: The efficacy of a commercial seed washer and 1 and 3% peroxyacetic acid or 20 000 ppm calcium hypochlorite for reducing Salmonella on alfalfa seeds was investigated. Methods and Results: Alfalfa seeds were inoculated with Salmonella Stanley to achieve c. 5 log CFU g^?1. Seeds were then treated with 1 or 3% peroxyacetic acid or 20 000 ppm calcium hypochlorite for 15 min in a commercial seed washer that uses air to enhance contact of the sanitizer with the seed. Experiments were also conducted using industry and laboratory methods. An c. 1‐log reduction in number of Salm. Stanley was demonstrated regardless of the chemical treatment or method of treatment. Although this 1‐log reduction was significant (P < 0·05), differences among the treatments were not significant. Treating the seed with 1 and 3% peroxyacetic acid resulted in similar Salm. Stanley reductions of 1·77 and 1·34 log, respectively, not being statistically significant (P > 0·05). Conclusions: These results suggest that under conditions tested, 1 or 3% peroxyacetic acid solutions are equally effective as 20 000 ppm of Ca(OCl)₂ in the reduction of Salm. Stanley on alfalfa seed when used in conjunction with a commercial seed washer. Significance and Impact of the Study: A 1% peroxyacetic acid solution could potentially be used in place of 20 000 ppm of Ca(OCl)₂ for treatment of seeds used for sprouting_. The commercial seed washer did not enhance removal of Salm. Stanley from alfalfa seeds, but did facilitate removal of excess soil from seeds.     

数字PCR在食源性致病菌检测中的应用进展

食源性致病菌是食品安全的重大隐患,对人类健康造成极大危害,因此亟待研究和建立精准有效的食源性致病菌检测方法。随着单分子检测技术的快速发展,数字PCR技术因其具有超高的灵敏度、稳定性和低试剂消耗等优点而被广泛应用于食源性致病菌的检测。主要介绍了数字PCR的基本原理及其研究进展,深入探讨了其在检测大肠杆菌（Escherichia coli）、沙门氏菌（Salmonella）、金黄色葡萄球菌（Staphylococcus aureus）、空肠弯曲菌（Campylobacter jejuni）、志贺氏菌（Shigella）、克罗诺杆菌（Cronobacter）、副溶血性弧菌（Vibrio parahaemolyticus）、单核细胞增生李斯特氏菌（Listeria monocytogenes）和蜡状芽孢杆菌（Bacillus cereus）中的应用,为今后该技术在食源性致病菌检测中的研究与应用提供一定的技术性参考。     

Biofilms as a microbial hazard in the food industry: A scoping review

Biofilms pose a serious public health hazard with a significant economic impact on the food industry. The present scoping review is designed to analyse the literature published during 2001–2020 on biofilm formation of microbes, their detection methods, and association with antimicrobial resistance (if any). The peer-reviewed articles retrieved from 04 electronic databases were assessed using PRISMA-ScR guidelines. From the 978 preliminary search results, a total of 88 publications were included in the study. On analysis, the commonly isolated pathogens were Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Escherichia coli, Bacillus spp., Vibrio spp., Campylobacter jejuni and Clostridium perfringens. The biofilm-forming ability of microbes was found to be influenced by various factors such as attachment surfaces, temperature, presence of other species, nutrient availability etc. A total of 18 studies characterized the biofilm-forming genes, particularly for S. aureus, Salmonella spp., and E. coli. In most studies, polystyrene plate and/or stainless-steel coupons were used for biofilm formation, and the detection was carried out by crystal violet assays and/or by plate counting method. The strain-specific significant differences in biofilm formation were observed in many studies, and few studies carried out analysis of multi-species biofilms. The association between biofilm formation and antimicrobial resistance was not clearly defined. Further, viable but non-culturable form of the foodborne pathogens is posing an unseen (by conventional cultivation techniques) but potent threat to the food safety. The present review recommends the need for carrying out systematic surveys and risk analysis of biofilms in food chain to highlight the evidence-based public health concerns, especially in regions where microbiological food hazards are quite prevalent.     

Bacteriophages: A weapon against mixed-species biofilms in the food processing environment

Mixed-species biofilms represent the most frequent actual lifestyles of microorganisms in food processing environments, and they are usually more resistant to control methods than single-species biofilms. The persistence of biofilms formed by foodborne pathogens is believed to cause serious human diseases. These challenges have encouraged researchers to search for novel, natural methods that are more effective towards mixed-species biofilms. Recently, the use of bacteriophages to control mixed-species biofilms have grown significantly in the food industry as an alternative to conventional methods. This review highlights a comprehensive introduction of mixed-species biofilms formed by foodborne pathogens and their enhanced resistance to anti-biofilm removal strategies. Additionally, several methods for controlling mixed-species biofilms briefly focused on applying bacteriophages in the food industry have also been discussed. This article concludes by suggesting that using bacteriophage, combined with other ‘green’ methods, could effectively control mixed-species biofilms in the food industry.     

Smart Amp快速检测技术及其应用

SmartAmp是一种新的DNA等温扩增技术,具有操作简单、快速、成本低、灵敏度高、特异性强、背景低等优点。目前已在临床基因多态性检测、感染性疾病诊断等方面初步应用,为临床快速诊断提供了帮助,在食品和环境中病原体检测领域也显示了极大的应用发展潜力。我们简要概述SmartAmp技术原理、特点及其应用前景。