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This detailed volume explores state-of-the-art methods for the identification, quantification, and characterization of microbial proteins. Split into five parts, the content addresses global sample preparation and protein enrichment, subcellular fractionation, protein quantification, analysis of post-translational protein modifications, as well as metaproteomics, a relatively new branch of microbial proteomics that investigates the proteins of all microbes comprising an environmental consortium. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Microbial Proteomics: Methods and Protocols serves as a valuable and stimulating source for all beginners and advanced researchers in the field of microbial proteomics and beyond. Chapter 18 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
The Use of Mass Spectrometry Technology (MALDI-TOF) in Clinical Microbiology presents the state-of the-art for MALDI-TOF mass spectrometry. It is a key reference defining how MALDI-TOF mass spectrometry is used in clinical settings as a diagnostic tool of microbial identification and characterization that is based on the detection of a mass of molecules. The book provides updated applications of MALDI-TOF techniques in clinical microbiology, presenting the latest information available on a technology that is now used for rapid microbial identification at relatively low cost, thus offering an alternative to conventional laboratory diagnosis and proteomic identification systems. Although the main use of the technology has, until now, been identification or typing of bacteria from a positive culture, applications in the field of virology, mycology, microbacteriology and resistances are opening up new opportunities. - Presents updated applications of MALDI-TOF techniques in clinical microbiology - Describes the use of mass spectrometry in the lab, the principles of the technology, preparation of samples, device calibration and maintenance, treatment of microorganisms, and quality control - Presents key information for researchers, including possible uses of the technology, differences between devices, how to interpret results, and future applications - Covers the topic in a systematic and comprehensive manner that is useful to both clinicians and researchers
New advances in proteomics, driven largely by developments in mass spectrometry, continue to reveal the complexity and diversity of pathogenic mechanisms among microbes that underpin infectious diseases. Therefore a new era in medical microbiology is demanding a rapid transition from current procedures to high throughput analytical systems for the diagnosis of microbial pathogens. This book covers the broad microbiological applications of proteomics and mass spectrometry. It is divided into six sections that follow the general progression in which most microbiology laboratories are approaching the subject –Transition, Tools, Preparation, Profiling by Patterns, Target Proteins, and Data Analysis.
This book highlights the triumph of MALDI-TOF mass spectrometry over the past decade and provides insight into new and expanding technologies through a comprehensive range of short chapters that enable the reader to gauge their current status and how they may progress over the next decade. This book serves as a platform to consolidate current strengths of the technology and highlight new frontiers in tandem MS/MS that are likely to eventually supersede MALDI-TOF MS. Chapters discuss: Challenges of Identifying Mycobacterium to the Species level Identification of Bacteroides and Other Clinically Relevant Anaerobes Identification of Species in Mixed Microbial Populations Detection of Resistance Mechanisms Proteomics as a biomarker discovery and validation platform Determination of Antimicrobial Resistance using Tandem Mass Spectrometry
In the last quarter century, advances in mass spectrometry (MS) have been at the forefront of efforts to map complex biological systems including the human metabolome, proteome, and microbiome. All of these developments have allowed MS to become a well-established molecular level technology for microorganism characterization. MS has demonstrated its considerable advantage as a rapid, accurate, and cost-effective method for microorganism identification, compared to conventional phenotypic techniques. In the last several years, applications of MS for microorganism characterization in research, clinical microbiology, counter-bioterrorism, food safety, and environmental monitoring have been documented in thousands of publications. Regulatory bodies in Europe, the US, and elsewhere have approved MS-based assays for infectious disease diagnostics. As of mid-2015, more than 3300 commercial MS systems for microorganism identification have been deployed worldwide in hospitals and clinical labs. While previous work has covered broader approaches in using MS to characterize microorganisms at the species level or above, this book focuses on strain-level and subtyping applications. In twelve individual chapters, innovators, leaders and practitioners in the field from around the world have contributed to a comprehensive overview of current and next-generation approaches for MS-based microbial characterization at the subspecies and strain levels. Chapters include up-to-date reference lists as well as web-links to databases, recommended software, and other useful tools. The emergence of new, antibiotic-resistant strains of human or animal pathogens is of extraordinary concern not only to the scientific and medical communities, but to the general public as well. Developments of novel MS-based assays for rapid identification of strains of antibiotic-resistant microorganisms are reviewed in the book as well. Microbiologists, bioanalytical scientists, infectious disease specialists, clinical laboratory and public health practitioners as well as researchers in universities, hospitals, government labs, and the pharmaceutical and biotechnology industries will find this book to be a timely and valuable resource.
A multidisciplinary approach to understanding the fundamentals of mass spectrometry for bacterial analysis From chemotaxonomy to characterization of targeted proteins, Identification of Microorganisms by Mass Spectrometry provides an overview of both well-established and cutting-edge mass spectrometry techniques for identifying microorganisms. A vital tool for microbiologists, health professionals, and analytical chemists, the text is designed to help scientists select the most effective techniques for use in biomedical, biochemical, pharmaceutical, and bioterror defense applications. Since microbiological applications of mass spectrometry require a basic understanding of both microbiology and analytical chemistry, the editors have incorporated material from both disciplines so that readers from either field will come to understand the necessary principles of the other. Featuring contributions from some of the most recognized experts in both fields, this volume provides specific examples of fundamental methods as well as approaches developed in the last decade, including: * Metastable atom bombardment pyrolysis mass spectrometry * Matrix-assisted laser desorption/ionization mass spectrometry (MALDI) * MALDI time-of-flight mass spectrometry (MALDI-TOF MS) of intact bacteria * High-resolution Fourier transform mass spectrometry (FTMS) * Electrospray ionization (ESI) mass spectrometry Identification of Microorganisms by Mass Spectrometry represents the most comprehensive and up-to-date work on the topic currently available. It is liberally illustrated with figures and tables and covers every aspect of spectrometric identification of microorganisms, including experimental procedures, various means of sample preparation, data analysis, and interpretation of complex mass spectral data.
Introduction to forensic proteomics -- A proteomics tutorial -- Proteomic sample preparation techniques : toward forensic proteomic applications -- NextGen serology : leveraging mass spectrometry for protein-based human body fluid identification -- Informatics approaches to forensic body fluid identification by proteomic mass spectrometry -- Fingermarks as a new proteomic specimen : state of the art and perspective of in situ proteomics -- Human identification using genetically variant peptides in biological forensic evidence -- Proteomics in the analysis of forensic, archaeological, and paleontological bone -- Proteomics for microbial forensics -- ISO 17025 accreditation of method-based mass spectrometry for bioforensic analyses -- Unambiguous identification of ricin and abrin with advanced mass spectrometric assays -- Challenges in the development of reference materials for protein toxins -- The statistical defensibility of forensic proteomics.
Biological Identification provides a detailed review of, and potential future developments in, the technologies available to counter the threats to life and health posed by natural pathogens, toxins, and bioterrorism agents. Biological identification systems must be fast, accurate, reliable, and easy to use. It is also important to employ the most suitable technology in dealing with any particular threat. This book covers the fundamentals of these vital systems and lays out possible advances in the technology. Part one covers the essentials of DNA and RNA sequencing for the identification of pathogens, including next generation sequencing (NGS), polymerase chain reaction (PCR) methods, isothermal amplification, and bead array technologies. Part two addresses a variety of approaches to making identification systems portable, tackling the special requirements of smaller, mobile systems in fluid movement, power usage, and sample preparation. Part three focuses on a range of optical methods and their advantages. Finally, part four describes a unique approach to sample preparation and a promising approach to identification using mass spectroscopy. Biological Identification is a useful resource for academics and engineers involved in the microelectronics and sensors industry, and for companies, medical organizations and military bodies looking for biodetection solutions. - Covers DNA sequencing of pathogens, lab-on-chip, and portable systems for biodetection and analysis - Provides an in-depth description of optical systems and explores sample preparation and mass spectrometry-based biological analysis
High-quality research articles on proteomic analyses of microbial pathogens, made available in a handy form. Containing proven, high-quality research articles selected from the popular PROTEOMICS journal, this is a current overview of the latest research into the proteomics analysis of microbial pathogens as well as several review articles.
This book covers the state-of-the-art of modern MALDI (matrix-assisted laser desorption/ionization) and its applications. New applications and improvements in the MALDI field such as biotyping, clinical diagnosis, forensic imaging, and ESI-like ion production are covered in detail. Additional topics include MS imaging, biotyping/speciation and large-scale, high-speed MS sample profiling, new methods based on MALDI or MALDI-like sample preparations, and the advantages of ESI to MALDI MS analysis. This is an ideal book for graduate students and researchers in the field of bioanalytical sciences. This book also: • Showcases new techniques and applications in MALDI MS • Demonstrates how MALDI is preferable to ESI (electrospray ionization) • Illustrates the pros and cons associated with biomarker discovery studies in clinical proteomics and the various application areas, such as cancer proteomics