A strategic plan to develop a matrix-assisted laser desorption/ionization with time-of-flight (MALDI-TOF/TOF) tandem mass spectrometry (MS) method in conjunction with a biomarker database for rapid microbial identification is described in this technical memorandum. The objective is to develop rapid, highly sensitive and definitive protocols for detection and identification of BW agents such as Bacillus species and demonstrate their - effectiveness. The protocols will be based on MALDI-TOF MS technology applicable to intact microorganisms. MALDI can directly analyze biological samples for biomarkers (proteins, lipids, etc.). Forensic biomarkers can be used to identify microorganisms at species and strain levels. In addition to method development, a new mass spectral biomarker database could be created and populated for selected species and strains, starting with selected Bacillus species. This technology would create a new area for comprehensive forensic analysis of biological materials that are not addressed by current methods (PCR and DNA sequencing, - and immunoassays). This memorandum contains a review of Bacillus species and current detection and identification methods, and a review of MALDI-TOF/TOF MS methods and biomarker databases.
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.
The Prokaryotes is a comprehensive, multi-authored, peer reviewed reference work on Bacteria and Achaea. This fourth edition of The Prokaryotes is organized to cover all taxonomic diversity, using the family level to delineate chapters. Different from other resources, this new Springer product includes not only taxonomy, but also prokaryotic biology and technology of taxa in a broad context. Technological aspects highlight the usefulness of prokaryotes in processes and products, including biocontrol agents and as genetics tools. The content of the expanded fourth edition is divided into two parts: Part 1 contains review chapters dealing with the most important general concepts in molecular, applied and general prokaryote biology; Part 2 describes the known properties of specific taxonomic groups. Two completely new sections have been added to Part 1: bacterial communities and human bacteriology. The bacterial communities section reflects the growing realization that studies on pure cultures of bacteria have led to an incomplete picture of the microbial world for two fundamental reasons: the vast majority of bacteria in soil, water and associated with biological tissues are currently not culturable, and that an understanding of microbial ecology requires knowledge on how different bacterial species interact with each other in their natural environment. The new section on human microbiology deals with bacteria associated with healthy humans and bacterial pathogenesis. Each of the major human diseases caused by bacteria is reviewed, from identifying the pathogens by classical clinical and non-culturing techniques to the biochemical mechanisms of the disease process. The 4th edition of The Prokaryotes is the most complete resource on the biology of prokaryotes. The following volumes are published consecutively within the 4th Edition: Prokaryotic Biology and Symbiotic Associations Prokaryotic Communities and Ecophysiology Prokaryotic Physiology and Biochemistry Applied Bacteriology and Biotechnology Human Microbiology Actinobacteria Firmicutes Alphaproteobacteria and Betaproteobacteria Gammaproteobacteria Deltaproteobacteria and Epsilonproteobacteria Other Major Lineages of Bacteria and the Archaea
Relying on practical examples from the authors’ experience, this book provides a thorough and modern approach to controlling and monitoring microbial contaminations during the manufacturing of non-sterile pharmaceuticals. Offers a comprehensive guidance for non-sterile pharmaceuticals microbiological QA/QC Presents the latest developments in both regulatory expectations and technical advancements Provides guidance on statistical tools for risk assessment and trending of microbiological data Describes strategy and practical examples from the authors’ experience in globalized pharmaceutical companies and expert networks Offers a comprehensive guidance for non-sterile pharmaceuticals microbiological QA/QC Presents the latest developments in both regulatory expectations and technical advancements Provides guidance on statistical tools for risk assessment and trending of microbiological data Describes strategy and practical examples from the authors’ experience in globalized pharmaceutical companies and expert networks
The threat of biological and chemical terrorism has driven the demand for timely techniques that can quickly detect the agent or agents used in an attack. The detection and/or prevention of these potential security threats provide significant scientific and technical challenges due to the combination of possible agents and modes of delivery available. This book will present a thorough look at the importance and technological challenges of mass spectrometry (MS) for the detection & identification of biological and chemical threats. This new contribution’s general aims are to draw the attention of recognized practitioners, experts and graduate students trying to grasp the latest MS developments in the cutting-edge fields of MS-biodefense technologies for the rapid/early/specific sensitive threat detection of pathogens, viruses, explosives, mycotoxins, chemical agents, and biological markers of xenobiotic chemicals.
Identifying pathogens in food quickly and accurately is one of the most important requirements in food processing. The ideal detection method needs to combine such qualities as sensitivity, specificity, speed and suitability for on-line applications. Detecting pathogens in food brings together a distinguished international team of contributors to review the latest techniques in microbiological analysis and how they can best be used to ensure food safety.Part one looks at general issues, beginning with a review of the role of microbiological analysis in food safety management. There are also chapters on the critical issues of what to sample and how samples should be prepared to make analysis effective, as well as how to validate individual detection techniques and assure the quality of analytical laboratories. Part two discusses the range of detection techniques now available, beginning with traditional culture methods. There are chapters on electrical methods, ATP bioluminescence, microscopy techniques and the wide range of immunological methods such as ELISAs. Two chapters look at the exciting developments in genetic techniques, the use of biosensors and applied systematics.Detecting pathogens in food is a standard reference for all those concerned in ensuring the safety of food. Reviews the latest techniques in microbiological analysis and how they can best be used to ensure food safety Examines the role of microbiological analysis in food safety management and discusses the range of detection techniques available Includes chapters on electrical methods, ATP bioluminescence, microscopy techniques and immunological methods such as ELISAs
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
Clinical microbiologists are engaged in the field of diagnostic microbiology to determine whether pathogenic microorganisms are present in clinical specimens collected from patients with suspected infections. If microorganisms are found, these are identified and susceptibility profiles, when indicated, are determined. During the past two decades, technical advances in the field of diagnostic microbiology have made constant and enormous progress in various areas, including bacteriology, mycology, mycobacteriology, parasitology, and virology. The diagnostic capabilities of modern clinical microbiology laboratories have improved rapidly and have expanded greatly due to a technological revolution in molecular aspects of microbiology and immunology. In particular, rapid techniques for nucleic acid amplification and characterization combined with automation and user-friendly software have significantly broadened the diagnostic arsenal for the clinical microbiologist. The conventional diagnostic model for clinical microbiology has been labor-intensive and frequently required days to weeks before test results were available. Moreover, due to the complexity and length of such testing, this service was usually directed at the hospitalized patient population. The physical structure of laboratories, staffing patterns, workflow, and turnaround time all have been influenced profoundly by these technical advances. Such changes will undoubtedly continue and lead the field of diagnostic microbiology inevitably to a truly modern discipline. Advanced Techniques in Diagnostic Microbiology provides a comprehensive and up-to-date description of advanced methods that have evolved for the diagnosis of infectious diseases in the routine clinical microbiology laboratory. The book is divided into two sections. The first techniques section covers the principles and characteristics of techniques ranging from rapid antigen testing, to advanced antibody detection, to in vitro nucleic acid amplification techniques, and to nucleic acid microarray and mass spectrometry. Sufficient space is assigned to cover different nucleic acid amplification formats that are currently being used widely in the diagnostic microbiology field. Within each technique, examples are given regarding its application in the diagnostic field. Commercial product information, if available, is introduced with commentary in each chapter. If several test formats are available for a technique, objective comparisons are given to illustrate the contrasts of their advantages and disadvantages. The second applications section provides practical examples of application of these advanced techniques in several "hot" spots in the diagnostic field. A diverse team of authors presents authoritative and comprehensive information on sequence-based bacterial identification, blood and blood product screening, molecular diagnosis of sexually transmitted diseases, advances in mycobacterial diagnosis, novel and rapid emerging microorganism detection and genotyping, and future directions in the diagnostic microbiology field. We hope our readers like this technique-based approach and your feedback is highly appreciated. We want to thank the authors who devoted their time and efforts to produce their chapters. We also thank the staff at Springer Press, especially Melissa Ramondetta, who initiated the whole project. Finally, we greatly appreciate the constant encouragement of our family members through this long effort. Without their unwavering faith and full support, we would never have had the courage to commence this project.
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.
