分子生物学技术应用于病原微生物分子分型的研究进展

病原微生物的分型检测在临床感染性疾病的诊断中发挥着重要作用。随着人类基因组计划的完成,分子生物学技术为病原微生物的分型检测提供了更多的分子靶点,进一步促进了病原微生物分型技术的发展,逐步取代了传统的分型方法。本文就近年来分子生物学技术应用于病原微生物分子分型的研究进行了综述。     

基因芯片在病原微生物检测中的应用评价

基因芯片技术是随着“人类基因组计划”的进展而发展起来的。至今 ,芯片技术已在医学领域中显示了其巨大的发展潜力并取得了一定的成功 ,为病原微生物的分子诊断技术提供了良好的发展前景。基因芯片技术的日臻完善 ,使基因芯片能同步进行微生物的诊断和测试抗菌药物的敏感性 ,实现病原微生物检测的技术革命。     

环介导等温扩增技术及其在分子诊断中的应用

环介导等温扩增技术作为新兴的分子生物学技术,已经广泛应用于各种病原微生物的分子水平的研究,并且取得了令人鼓舞的成绩。文中就环介导等温扩增技术的原理及在分子诊断中的应用作一综述。     

拉曼光谱技术在检验医学中的研究进展及应用

拉曼光谱技术以其“指纹图谱”性、无损检测、灵敏度高、简便快速等独特的技术优势, 在生物大分子、病原微生物检测、肿瘤精准分子诊断等领域展现出良好的应用潜力。该文立足于临床检验应用,简要阐述了拉曼光谱检测的技术原理,从生物大分子、细胞、组织、病原微生物、体液检测等多层次详细介绍了拉曼光谱技术在生物医学应用方面的技术发展及最...     

基因芯片在病原微生物检测中的应用评价

基因芯片技术是随着“人类基因组计划”的进展而发展起来的。至今，芯片技术已在医学领域中显示了其巨大的发展潜力并取得了一定的成功，为病原微生物的分子诊断技术提供了良好的发展前景。基因芯片技术的日臻完善，使基因芯片能同步进行微生物的诊断和测试抗菌药物的敏感性，实现病原微生物检测的技术革命。     

血流感染病原微生物诊断技术的研究与应用

血流感染(BSI)是指病原微生物侵入血液循环并在体内繁殖的一种全身感染性疾病。近年来因其高发病率及病死率而受到越来越多的关注,如何实现BSI致病菌的快速检测及其药敏结果的准确回报,是长期困扰临床的问题。血培养是诊断BSI病原微生物的"金标准",但其阳性率低且结果报告周期长,因此需探索新的技术以弥补此缺陷。近年来,微生物多种诊断技术,如阳性报警时间、核酸杂交技术、核酸扩增技术、质谱检测技术、基因芯片和测序技术等逐步从研究阶段走向BSI病原微生物诊断的临床应用阶段。本文旨在对BSI病原微生物基于全血标本和基于血培养的检测技术进行综述,为BSI病原微生物快速诊断和治疗提供方法学依据。     

环介导恒温扩增技术在百日咳鲍特菌检测中应用研究进展

正核酸扩增技术是病原微生物检测的重要武器,其中聚合酶链反应(PCR)技术已经在临床检验中广泛应用。与PCR相比,环介导恒温扩增(LAMP)技术具有操作简便,快速灵敏,对仪器和样本质量要求低等优点,尤其适合病原微生物的即时检验。本文主要综述了LAMP技术在百日咳即时检验中应用的研究     

环介导等温扩增技术在病原微生物临床检测中的应用

病原微生物是威胁人类健康的重要因素之一,快速、准确的检测方法对于感染性疾病的诊断具有重要意义。环介导等温扩增(LAMP)是一种恒温扩增方法,具有反应时间短、操作简便、灵敏度高、特异度高、检测成本低等优点,因此被广泛应用于感染性疾病的快速诊断。基于这种情况,该文主要阐述了LAMP的原理、特点及其在临床常见病原微生物检测方面的应用与研究进展。LAMP技术现在已经被应用于病原微生物的检测,且具有较高的灵敏度及特异度,但该技术仍有易产生假阳性、引物设计复杂等不足之处。该文综述了近年来LAMP技术在病原微生物感染中的应用与进展,并对其未来的发展前景进行了展望,以期在资源有限的环境中为病原微生物感染的快速诊断提供合理的研究方向。     

高通量测序在病原微生物领域应用进展

目前高通量测序是指一次性对上百万乃至上亿条DNA分子进行的并行测序技术。高通量测序使人类能够以短时间、低成本的方式从基因组学角度对各种生命现象做出精细描绘,同时该技术也为后基因组时代,即蛋白质组学的深入研究提供了准确蓝本。本文就高通量测序在病原微生物领域的应用进展进行总结。从病原微生物分类、传染病监测、病原微生物耐药以及人类微生物组分析这四方面进行综述。     

微生物在线生物信息分析系统的开发及应用

目的 基于浏览器/服务器模式构建微生物在线生物信息分析系统,应用于传染病预防控制工作。 方法 将目前在传染病病原微生物研究和传染病分子诊断、溯源中常用的基于客户端/服务器模式的一些开源的或免费提供使用的分析软件,采用远程桌面、分布式处理等"云计算"技术方案,构建成不需要在本地安装,以浏览器/服务器模式直接通过浏览器以点鼠标的方式使用的微生物在线生物信息分析系统。 结果 构建了包含引物设计、序列比对和进化树构建、数据统计、图形展示四类共15种常用生物信息分析功能的微生物在线生物信息分析系统,实现了通过浏览器进行病原微生物序列数据在线分析及疫情溯源。 结论 通过构建基于浏览器/服务器模式的微生物在线生物信息分析系统,可为疾病预防控制工作者提供方便快捷的生物信息分析服务,加快病原微生物序列数据的深度挖掘和疫情溯源,提高应对突发疫情的反应速度。     

Dual-modality optical coherence tomography and fluorescence tethered capsule endomicroscopy

OCT tethered capsule endomicroscopy (TCE) is an emerging noninvasive diagnostic imaging technology for gastrointestinal (GI) tract disorders. OCT measures tissue reflectivity that provides morphologic image contrast, and thus is incapable of ascertaining molecular information that can be useful for improving diagnostic accuracy. Here, we introduce an extension to OCT TCE that includes a fluorescence (FL) imaging channel for attaining complementary, co-registered molecular contrast. We present the development of an OCT-FL TCE capsule and a portable, plug-and-play OCT-FL imaging system. The technology is validated in phantom experiments and feasibility is demonstrated in a methylene blue (MB)-stained swine esophageal injury model, ex vivo and in vivo.     

The value of portable ultrasound for evaluation of cardiomegaly patients presenting at the emergency department

OBJECTIVES: The purpose of this study was to evaluate the value of a hand-carried portable ultrasound device in the emergency department for patients with cardiomegaly and evaluate its impact on diagnosis and treatment. We compared the results of the portable ultrasound with the results of physical examination and with standard echocardiograph (SE) in the cardiomegaly patients in emergency department. BACKGROUND: Recently, small portable ultrasound devices have been introduced, and they need more extensive study to evaluate their application. METHODS: We used the OptiGo (Agilent Technologies, Andover, MA) portable device to evaluate emergency patients, and compared it with two SE devices available for this study, the Hewlett Packard (Sono 5500; Andover, MA) or the Vingmed (System V; Horten, Norway). Each of 100 patients was studied by physical examination first then examined with the portable ultrasound and standard echocardiography. The yields from physical examination and portable echocardiography were compared and results of the standard and portable were also compared. RESULTS: There were a total of 243 cardiovascular findings detected by the standard echocardiographic examination in the 100 patients studied. Cardiac examination failed to detect 40% of the overall findings but the portable device missed only 17% of all findings. The portable device evaluated 201 conditions correctly (83%) and missed relevant clinical findings in 37 (17%) as compared to standard echocardiography, but in only 12 (5%) were these findings of major importance As an overall measure of diagnostic value, the portable device would have added to clinical judgment, thus boosting diagnostic accuracy from 62% to 83%. CONCLUSIONS: Portable ultrasound technology can provide rapid, readily available and important clinical information for emergency physicians in the management of emergency patients with cardiomegaly.     

Bedside blood gas and electrolyte monitoring in critically ill patients

A major advantage of near-patient testing is time savings that facilitate important diagnostic and therapeutic decisions. Recent technologic advances have made available a number of systems that allow for near-patient testing. The reliability of these instruments must be validated in the clinical setting in the hands of their intended users. We evaluated the Gemstat blood gas, electrolyte, and Hct portable analyzer in the critical care setting when used by numerous individuals with no previous laboratory training. Blood gas, Na, K, and Hct results were highly correlated with those from the clinical laboratories (PaO2, r = .96; PaCO2, r = .92, pH, r = .96; Na, r = .93; K, r = .95; Hct, r = .91). The Gemstat represents a new generation of portable, rapid, safe, and accurate instruments that are well suited for ICU settings. The instrument can facilitate clinical management of patients, and may improve patient care.     

Portable Ultrasound for Remote Environments,Part II: Current Indications

Background: With recent advances in ultrasound technology, it is now possible to deploy lightweight portable imaging devices in the field. Techniques and studies initially developed for hospital use have been extrapolated out of the hospital setting in a wide variety of environments in an effort to increase diagnostic accuracy in austere or prehospital environments. Objectives: This review summarizes current ultrasound applications used in out-of-hospital arenas and highlights existing evidence for such use. The diversity of applications and environments is organized by indication to better inform equipment selection as well as future directions for research and development. Discussion: Trauma evaluation, casualty triage, and assessment for pneumothorax, acute mountain sickness, and other applications have been studied by field medical teams. A wide range of outcomes have been reported, from alterations in patient care to determinations of accuracy compared to clinical judgment or other diagnostic modalities. Conclusions: The use of lightweight portable ultrasound shows great promise in augmenting clinical assessment for field medical operations. Although some studies of diagnostic accuracy exist in this setting, further research focused on clinically relevant outcomes data is needed.     

基于CRISPR/Cas系统诊断平台的研究进展

目前核酸检测技术已被广泛应用于临床实验室诊断,常规检测技术如实时荧光定量PCR技术耗时长且依赖特定的仪器设备。成簇规律间隔短回文重复序列(CRISPR)/CRISPR相关蛋白(Cas)系统是细菌和古细菌在与病毒斗争过程中获得的适应性免疫防御机制,已被发展成强大的基因组编辑技术。最近CRISPR领域的先驱团队基于Cas13a、Cas12a和新发现的Cas14蛋白开发出SHERLOCK、DETECTR等新型核酸检测工具,在传染性疾病的快速诊断、癌症中基因突变的检测和基因分型等方面意义重大,其灵敏度高、特异性强且快速经济,在临床分子诊断领域具有巨大潜力。本文综述了CRISPR/Cas系统的作用机制及新型诊断平台的原理和应用进展,总结了新型检测技术的优缺点并对其发展前景进行展望。     

BioMEMS using electrophoresis for the analysis of genetic mutations

Biomedical microelectromechanical systems (BioMEMS) are rapidly emerging in many areas of genetic analysis. These devices demonstrate potential for rapid analysis using modular components capable of sample purification, amplification, mutation discrimination and detection on small, portable point-of-care instruments. Here, various approaches to genetic mutation detection and the modern analysis platform, capillary electrophoresis, will be briefly reviewed. Microfluidic devices will be discussed in relation to fabrication techniques, mutation detection using simple electrophoretic separations, multiplexed designs and modular functionalities, as well as challenges and issues surrounding this technology.     

Non-culture based diagnostics for intravascular catheter related bloodstream infections

Introduction: intravascular catheter related bloodstream infection (IVC-BSI) is a leading cause of nosocomial infections and associated with significant morbidity and mortality. Early detection and adequate treatment of causative pathogens is critical for a favourable outcome. However, it takes significant time to receive microbiological results due to the current reference diagnostic method’s reliance on microbial growth.

Areas covered: This review discusses culture and non-culture based techniques for the diagnosis of non IVC-BSI and IVC-BSI, including molecular methods and biomarkers. Different diagnostic strategies are evaluated and the potential of new generation of diagnostic assays highlighted.

Expert commentary: The development of additional diagnostic methods has potential to beneficially supplement conventional culture diagnosis, and molecular techniques have particular potential to fulfil this need. They would also contribute significant new knowledge on the bacterial species present on catheters that are generally missed by diagnosis using traditionally culture-dependent methods. Advances in molecular strategies, together with new biomarkers, might lead to the development of faster, more sensitive and cheaper technologies and instruments. This review aims to provide a platform for the further development of IVCBSI diagnostic techniques.     

Portable devices and mobile instruments for infectious diseases point-of-care testing

Introduction: Rapidity, simplicity, and portability are highly desirable characteristics of tests and devices designed for performing diagnostics at the point of care (POC), either near patients managed in healthcare facilities or to offer bioanalytical alternatives in external settings. By reducing the turnaround time of the diagnostic cycle, POC diagnostics can reduce the dissemination, morbidity, and mortality of infectious diseases and provide tools to control the global threat of antimicrobial resistance.

Areas covered: A literature search of PubMed and Google Scholar, and extensive mining of specialized publications, Internet resources, and manufacturers’ websites have been used to organize and write this overview of the challenges and requirements associated with the development of portable sample-to-answer diagnostics, and showcase relevant examples of handheld devices, portable instruments, and less mobile systems which may or could be operated at POC.

Expert commentary: Rapid (<1 h) diagnostics can contribute to control infectious diseases and antimicrobial resistant pathogens. Portable devices or instruments enabling sample-to-answer bioanalysis can provide rapid, robust, and reproducible testing at the POC or close from it. Beyond testing, to realize some promises of personalized/precision medicine, it will be critical to connect instruments to healthcare data management systems, to efficiently link decentralized testing results to the electronic medical record of patients.     

Current applications of duplex and color Doppler ultrasound imaging: carotid and peripheral vascular system

Recent advances in ultrasound technology have made possible the development of diagnostic instruments that combine cross-sectional imaging and Doppler analysis. These instruments have expanded the role of diagnostic ultrasonography to the assessment of carotid and peripheral vascular disease. The current applications of duplex Doppler and color Doppler imaging in evaluating the extracranial carotid arteries, vertebral artery, peripheral venous system, and peripheral arterial system are reviewed. The indications for and limitations of these examinations, as well as the potential future uses, are discussed.     

Novel droplet platforms for the detection of disease biomarkers

Personalized medicine – healthcare based on individual genetic variation – has the potential to transform the way healthcare is delivered to patients. The promise of personalized medicine has been predicated on the predictive and diagnostic power of genomic and proteomic biomarkers. Biomarker screening may help improve health outcomes, for example, by identifying individuals’ susceptibility to diseases and predicting how patients will respond to drugs. Microfluidic droplet technology offers an exciting opportunity to revolutionize the accessibility of personalized medicine. A framework for the role of droplet microfluidics in biomarker detection can be based on two main themes. Emulsion-based microdroplet platforms can provide new ways to measure and detect biomolecules. In addition, microdroplet platforms facilitate high-throughput screening of biomarkers. Meanwhile, surface-based droplet platforms provide an opportunity to develop miniaturized diagnostic systems. These platforms may function as portable benchtop environments that dramatically shorten the transition of a benchtop assay into a point-of-care format.