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Development and evaluation of a multiplex PCR for simultaneous detection of five foodborne pathogens 总被引:2,自引:0,他引:2
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. 相似文献
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E. Patrick Groody 《Molecular biotechnology》1996,6(3):323-327
The detection of foodborne microorganisms has traditionally been done using microbiologically based methods. Such “gold standard”
methods are generally reliable but have the disadvantages of being labor intensive, subjective, and time consuming. Over the
last several years, the development of DNA probe-based methods has simplified the methods used to detect organisms such asSalmonella, Listeria, andE. coli by targeting the unique DNA or RNA sequences of these organisms using DNA probes and nonradioactive detection. 相似文献
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摘要:贝类是食源性致病微生物的重要传播载体之一,因食用贝类导致食源性疾病的发病率逐年升高。因此,贝类食用安全监测与控制成为食品安全研究的重点。近几年,笔者在贝类中主要致病微生物分子检测、累积分布和控制等方面开展了一系列研究。本文结合国内外的研究进展,对贝类中致病微生物的检测方法、组织分布、净化以及流行病学等方面展开论述。综合国内外研究结论发现,分子检测方法已成为贝类中致病微生物检测的主要手段。另外,贝类中致病微生物主要累积富集在鳃组织和消化腺(包括肠胃和消化盲囊)中,这为致病微生物的检测靶向性提供了理论指导。 相似文献
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现代分子生物学技术在瘤胃微生态系统研究中的应用 总被引:4,自引:0,他引:4
瘤胃中栖息着大量的微生物,由于这些微生物组成复杂且有些细菌在体外无法培养,目前对这些微生物的了解仍然很少。现代分子生物学技术的发展为研究瘤胃微生物提供了有效的方法,利用核酸探针、基因序列分析、遗传指纹技术、全细胞杂交和实时定量PCR等技术可以对瘤胃微生物的分类及进化关系、区系结构图、重要酶的表达以及目的微生物的准确定量进行更为深入和透彻的研究。发展和利用这些技术不仅可以研究微生物之间的关系以及微生物与饲料颗粒之间时间与空间的关系,还能直接在细菌自然生长的环境中对其各种特征进行研究。 相似文献
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Mariachiara Conte Francesco Aliberti Laura Fucci Marina Piscopo 《World journal of microbiology & biotechnology》2007,23(12):1679-1683
Antibacterial effects of various arginine- and lysine-rich polycationic proteins and polymers were evaluated by broth and
solid dilution assay on a range of foodborne pathogens, Gram-positive and Gram-negative bacteria. The Minimum Inhibitory Concentration
(MIC) and the Minimum Bactericidal Concentration (MBC) of α-poly-l-lysine (poly-lys), α-poly-l-arginine (poly-arg) and protamines from herring sperm (clupeine sulphate) and salmon sperm (salmine sulphate) were determined
on Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium, Shigella sonnei,
Escherichia coli O157:H7 and Pseudomonas aeruginosa. All these molecules showed antibacterial activity on all strains with different MIC and MBC values. The molecular mechanisms
underlying the effect of α-poly-l-arginine might be related to the entrance of the molecule into the cell. In fact α-poly-l-arginine labelled with 7-Diethylamino coumarin-3-carboxylic acid, succinimidyl ester (DEAC,SE) showed ability to permeate
the cell membrane of B. cereus and E. coli O157:H7. 相似文献
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Zijie Chen Zhen Liu Jingjing Liu Xilin Xiao 《Biotechnology and bioengineering》2023,120(12):3501-3517
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. 相似文献
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肽核酸在分子生物学技术中的应用 总被引:1,自引:0,他引:1
肽核酸(PNA)作为一种人工合成的核酸类似物,以中性的肽链酰胺2-氨基乙基甘氨酸键取代了DNA中的戊糖磷酸二酯键骨架,其余部分与DNA相同。PNA可通过Watson-Crick碱基配对的形式识别并结合DNA或RNA序列,形成稳定的双螺旋结构。与传统的DNA或RNA相比,PNA具有生物学稳定性高、杂交特异性强、杂合体的稳定性高和杂交速度快等明显优点,使PNA具有良好的物理化学性质和生物学特性,在检测目的核酸序列中单碱基突变、PCR基因分子诊断与检测、荧光原位杂交定量分析、基因芯片和生物传感器技术等调控水平和临床应用上有自己的特点。简要综述了近年来肽核酸在上述分子生物学技术中的运用以及应用前景的展望。 相似文献
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Microfluidic devices for sample preparation and rapid detection of foodborne pathogens 总被引:1,自引:0,他引:1
Krishna Kant Mohammad-Ali Shahbazi Vivek Priy Dave Tien Anh Ngo Vinayaka Aaydha Chidambara Linh Quyen Than Dang Duong Bang Anders Wolff 《Biotechnology advances》2018,36(4):1003-1024
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. 相似文献
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结核病一直是世界性问题,我国其发病情况尤为严重,是亚洲的第二大结核病发病国家。结核病治疗方面常使用抗生素作为首选药物,随着抗菌药的滥用,结核杆菌对多种抗菌药产生耐药性,结核病耐药患者增多,治疗难度增加。因此,结核杆菌耐药分子机制的研究更加重要,新型抗结核药物研制更加迫切。结核分枝杆菌的基因突变是引起耐药的主要分子学依据,因此基于结核分枝杆菌耐药性相关基因的深入探索,对于预防结核病的传播及治疗皆具有深远影响。本文从分子生物学角度分析了近年来结核分枝杆菌耐药性产生的原因及相关研究进展。 相似文献
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人类免疫缺陷病毒(HIV)是获得性免疫缺陷综合征(AIDS)的病原体。根据血清学反应和核酸序列测定将HIV分为HIV-1和HIV-2两型,本研究就HIV-1基因的分子特征、HIV基因分型分类、HIV基因分型常用的方法、HIV-1病毒载量的分子生物学检测方法等方面进行讨论。 相似文献