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。     

生物传感器应用于食源性致病菌检测研究进展

生物传感器技术是一种由生物、化学、物理、医学、电子技术等多种学科互相渗透形成起来的高新微量分析技术,具有选择性好、灵敏度高、分析速度快、成本低、能在复杂的体系中进行在线连续监测的特点.本文根据生物传感器的分子识别元件将生物传感器分为DNA传感器、免疫传感器、细胞传感器三大类,简要介绍各种生物传感器的原理及其在检测食源性致病菌方面的应用情况,并对未来生物传感器应用于实际检测进行了展望.     

食源性致病菌快速检测技术研究进展

食源性致病菌是影响食品安全的主要因素之一,传统的细菌分离、培养与鉴定由于需时较长,特别是有的细菌难以培养,难以适应食源性疾病预防控制的需要,因而快速、简便、特异的检测方法成为研究的热点。对电阻抗、放射测量、微热量、ELISA、PCR、基因芯片和生物传感器技术在金黄色葡萄球菌、沙门菌、肠出血性大肠埃希菌等食源性致病菌快速检测中的应用研究进行综述。     

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。     

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。     

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。     

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。     

检测食源性致病菌的生物传感器

  大肠埃希氏菌、李斯特氏菌和鼠伤寒沙门氏菌等几种食源性致病菌不仅威胁到人们的 生命安全,还会造成巨大的社会经济损失.生物传感器是将生物识别元件和信号转换元件紧 密结合,从而检测目标化合物的分析装置.生物传感器在致病菌检测方面具有分析速度快、 灵敏度高、专一性强等特点;可分为光学式、电化学式、压电式生物传感器等;在检测食源 性致病菌方面生物传感器表现出能够满足实际应用的发展潜力,但是生物传感器目前仍面临 并需要解决一些问题,这也是生物传感器从实验室到市场如此缓慢的原因.最后提出了实际 检测应用中对生物传感器的要求.     

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。     

生物传感器在食源性致病菌大肠杆菌O157∶H7检测中的应用进展

食源性致病菌是引发食物中毒的主要微生物，对人类的健康构成了重要威胁，也逐渐发展为公共卫生、食品工业部门高度重视的一个问题。近年来，因食源性致病菌产生的疾病在世界各地蔓延，其中，大肠杆菌O157∶H7是食源性致病菌种类中造成病死率较高的一种细菌，其感染剂量低、致病性强等特点引起研究人员和世界各国卫生组织的广泛关注。基于此，对大肠杆菌O157∶H7的检测方法及其演变进行了梳理和比较分析，并着重综述了生物传感器（biosensor）在其检测中的应用进展，以期为大肠杆菌O157∶H7快速、准确和可商业化的检测方法的研发提供参考。     

食源性致病菌群体感应信号分子的检测

群体感应(Quorum sensing,QS)在食物中毒导致的食源性疾病暴发机制和食物腐败变质中起主要作用,QS影响致病菌的细胞被膜形成和致病性。文中通过深入了解食源性致病菌的QS信号分子,综述了革兰氏阴性和革兰氏阳性菌产生的信号分子类型,同时介绍了检测QS信号分子的不同技术,并根据QS机制在食品中的影响提出了思考和建议,为监控食源性致病菌提供依据。     

Development and evaluation of a multiplex PCR for simultaneous detection of five foodborne pathogens

Aims: To develop a rapid multiplex PCR method for simultaneous detection of five major foodborne pathogens (Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, Salmonella Enteritidis and Shigella flexneri, respectively). Methods and Results: Amplification by PCR was optimized to obtain high efficiency. Sensitivity and specificity assays were investigated by testing different strains. With a multipathogen enrichment, multiplex PCR assay was able to simultaneously detect all of the five organisms in artificially contaminated pork samples. The developed method was further applied to retail meat samples, of which 80% were found to be positive for one or more of these five organisms. All the samples were confirmed by traditional culture methods for each individual species. Conclusions: This study reported a rapid multiplex PCR assay using five primers sets for detection of multiple pathogens. Higher consistency was obtained between the results of multiplex PCR and traditional culture methods. Significance and Impact of the Study: This work has developed a reliable, useful and cost‐effective multiplex PCR method. The assay performed equally as well as the traditional cultural method and facilitated the sensitive detection both in artificially contaminated and naturally contaminated samples.     

Research progress in the detection of common foodborne hazardous substances based on functional nucleic acids biosensors

With the further improvement of food safety requirements, the development of fast, highly sensitive, and portable methods for the determination of foodborne hazardous substances has become a new trend in the food industry. In recent years, biosensors and platforms based on functional nucleic acids, along with a range of signal amplification devices and methods, have been established to enable rapid and sensitive determination of specific substances in samples, opening up a new avenue of analysis and detection. In this paper, functional nucleic acid types including aptamers, deoxyribozymes, and G-quadruplexes which are commonly used in the detection of food source pollutants are introduced. Signal amplification elements include quantum dots, noble metal nanoparticles, magnetic nanoparticles, DNA walkers, and DNA logic gates. Signal amplification technologies including nucleic acid isothermal amplification, hybridization chain reaction, catalytic hairpin assembly, biological barcodes, and microfluidic system are combined with functional nucleic acids sensors and applied to the detection of many foodborne hazardous substances, such as foodborne pathogens, mycotoxins, residual antibiotics, residual pesticides, industrial pollutants, heavy metals, and allergens. Finally, the potential opportunities and broad prospects of functional nucleic acids biosensors in the field of food analysis are discussed.     

Microfluidic devices for sample preparation and rapid detection of foodborne pathogens

Rapid detection of foodborne pathogens at an early stage is imperative for preventing the outbreak of foodborne diseases, known as serious threats to human health. Conventional bacterial culturing methods for foodborne pathogen detection are time consuming, laborious, and with poor pathogen diagnosis competences. This has prompted researchers to call the current status of detection approaches into question and leverage new technologies for superior pathogen sensing outcomes. Novel strategies mainly rely on incorporating all the steps from sample preparation to detection in miniaturized devices for online monitoring of pathogens with high accuracy and sensitivity in a time-saving and cost effective manner. Lab on chip is a blooming area in diagnosis, which exploits different mechanical and biological techniques to detect very low concentrations of pathogens in food samples. This is achieved through streamlining the sample handling and concentrating procedures, which will subsequently reduce human errors and enhance the accuracy of the sensing methods. Integration of sample preparation techniques into these devices can effectively minimize the impact of complex food matrix on pathogen diagnosis and improve the limit of detections. Integration of pathogen capturing bio-receptors on microfluidic devices is a crucial step, which can facilitate recognition abilities in harsh chemical and physical conditions, offering a great commercial benefit to the food-manufacturing sector. This article reviews recent advances in current state-of-the-art of sample preparation and concentration from food matrices with focus on bacterial capturing methods and sensing technologies, along with their advantages and limitations when integrated into microfluidic devices for online rapid detection of pathogens in foods and food production line.     

人工模拟胃肠道模型在食源性致病菌耐受及致病机理中的应用

人工模拟胃肠道模型是研究食源性致病菌耐受及致病机理的一种重要工具,其本质是在实验室模拟的条件下,重现人体消化过程中的化学、物理及生物作用,以研究食源性致病菌的耐受性、致病机理、肠道菌群互作及疫苗开发,对食源性致病菌的控制和治疗具有十分重要的意义。文中综述了人工模拟胃肠道模型在食源性致病菌研究中的应用,将现有胃肠道模型系统地划分为体外静态模型、体外动态模型、普通动物模型及人源化动物模型,并详细介绍了各类模型的概念及特性。在此基础之上,进一步分析了现有模型的不足,并对未来人工模拟胃肠道模型的研究方向进行了展望,以期为食源性致病菌耐受及致病机理的研究奠定扎实的研究基础。     

Application of antimicrobial ice for reduction of foodborne pathogens (Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes) on the surface of fish

AIMS: The efficacy of antimicrobial ice was evaluated for the reduction of foodborne pathogens on the surface of fish. METHODS AND RESULTS: Antimicrobial ice containing chlorine dioxide (ClO2) was utilized to control foodborne pathogens in laboratory media and on fish skin. Escherichia coli O157:H7, Salmonella serotype Typhimurium and Listeria monocytogenes strains were treated with antimicrobial ice for 30 min on plates of selective agar and for 120 min on fish skin at room temperature, and then incubated for enumeration. After treatment with 100 ppm ClO2 for 30 min, 5.4, 4.4 and 3.2 log10 reduction was obtained with E. coli O157:H7, Salm. Typhimurium and L. monocytogenes on laboratory media, respectively. When antimicrobial ice (100 ppm ClO2) was applied to fish skin for 120 min, total reduction of E. coli O157:H7, Salm. Typhimurium and L. monocytogenes was 4.8, 2.6 and 3.3 log10, respectively. CONCLUSION: The initial load of foodborne pathogens was reduced by antimicrobial ice and the lowered microbial level was maintained during treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: The application of antimicrobial ice is a simple and effective method for the safe preservation of fish.     

Impact of molecular biology on the detection of foodborne pathogens

Molecular biological methods that use antibodies and nucleic acids to detect specific foodborne bacterial pathogens were scarcely known a decade and a half ago. Few scientists could have predicted that these tools of basic research would come to dominate the field of food diagnostics. Today, a large number of cleverly designed assay formats using these technologies are available commercially for the detection in foods of practically all major established pathogens and toxins, as well as of many emerging pathogens. These tests range from very simple antibody-bound latex agglutination assays to very sophisticated DNA amplification methods. Although molecular biological assays are more specific, sensitive, and faster than conventional (often cultural) microbiological methods, the complexities of food matrices continue to offer unique challenges that may preclude the direct application of these molecular biological methods. Consequently, a short cultural enrichment period is still required for food samples prior to analysis with these assays. The greater detection sensitivity of molecular biological methods may also affect existing microbiological specifications for foods; this undoubtedly will have repercussions on the regulatory agencies, food manufacturers, and also consumers. The US government has the right to retain a nonexclusive royalty-free license in and to any copyright covering this article. Use of trade names is for identification only and does not imply an endorsement by the US FDA.     

Conductometric immunosensors for the detection of staphylococcal enterotoxin B based bio-electrocalytic reaction on micro-comb electrodes

Staphylococcal enterotoxin B (SEB) is one of many toxins produced by the Gram-positive bacterium Staphylococcal aureus. While SEB is known as the causative agent of certain food poisonings it is also considered abiological select agent. Thus, rapid and accurate identification of SEB during either surveillance or in response to a biothreat is critical to the mitigation of the suspect agent. This report presents a new conductometric immune-biosensor for the detection of SEB based on immobilization of horseradish peroxidase (HRP)-labeled SEB antibody (HRP-anti-SEB) onto nanogold/chitosan-multiwalled carbon nanotube (Au/CTS-MWNT)-functionalized biorecognition interface. The formation of the antibody-antigen complex by a simple one-step immunoreaction between the immobilized HRP-anti-SEB and SEB in sample solution introduced a barrier of electrical communication between the immobilized HRP and the base surface, thus local conductivity variations could be evaluated by the bio-electrocatalytic reaction of HRP in 0.02 M PBS (pH 6.8) containing 0.15 mM H(2)O(2), 0.06 M KI and 0.1 M NaCl. Under optimal conditions, the proposed immune-biosensor exhibited a good conductometric response relative to SEB concentration in a linear range from 0.5 to 83.5 ng/ml with a correlation coefficient of 0.998. The developed immune-biosensor showed an acceptable accuracy, reproducibility and stability. Milk samples spiked with various concentrations of SEB gave an average of 116% recovery of the toxin.     

病原中的活性氧释放研究进展

活性氧的释放在动植物-病原菌互作过程中有着非常重要的作用.一般认为互作中的活性氧来源于动植物细胞生物膜中的氧化还原体系.但近年来随着互作研究的深入,发现动植物病原菌自身也有活性氧的释放以及复杂的调控系统,它们的活性氧释放能力很有可能与其致病性有一定的联系,并可能参与了互作,这些发现对深入了解动植物-病原菌的互作机制具有重要意义.概述了在细菌、真菌等多种动物病原菌中存在的活性氧释放现象,这些微生物活性氧产生的位点、相关功能分子以及调控机制,介绍了目前研究仍然较少但其潜在意义重大的植物病原菌中的活性氧释放现象、可能的调节机制和病理学意义.