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Hands-on laboratory experts present a set of "classic" PCR-based methods for the identification and detection of important animal and food microbial pathogens, including several zoonotic agents. These proven techniques can be precisely applied to a wide variety of microbes, among them Campylobacter spp., chlamydiae, toxigenic clostridia, Escherichia coli (STEC), Listeria monocytogenes, mycoplasmas, salmonellae, and Yersinia enterocolitica. Additional chapters review the specificity and performance of diagnostic PCR analysis, the pre-PCR processing of samples, the critical aspects of standardizing PCR methods, and the general issues involved in using PCR technology for microbial diagnosis.
PCR methods for the detection of microbial pathogens have made relatively little impact in diagnostic microbiology laboratories due to the common decision to use expensive commercially produced tests rather than the cheaper alternative of developing one’s own tests or introducing tests developed by other workers. PCR Detection of Microbial Pathogens, Second Edition presents alternatives to commercially produced PCR methods to detect microbial pathogens. Although most of the chapters in this book are devoted to the detection of specific pathogens, the first chapters in this book should appeal to anyone working in this field regardless of their particular interests. Although PCR tests can often be made to work with relatively little effort, it is often unclear how efficient the PCR test is, how inhibitory the specimen containing the pathogen of interest is and how the test can be quality controlled. All of which are of great importance in developing tests for diagnostic use. These topics are covered in great depth at the beginning of the book. The main part of the book is devoted to describing methods for the detection of a wide range of pathogens and from widely different specimens and situations. Written in the highly successful Methods in Molecular BiologyTM series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, PCR Detection of Microbial Pathogens, Second Edition serves microbiologists regardless of their particular interest because, when used together with the general principles, the sheer variety of procedures provided here enables the reader to design and introduce diagnostic tests in the laboratory with confidence.
Not another textbook, but a valuable tool for doctors and microbiologists wanting to know how to set up a PCR diagnostic microbiology laboratory according to current regulatory standards and perform assays supplied with patient clinical diagnostic criteria and easy to follow protocols. Whether laboratories are using commercial kits or in-house methods developed in their own laboratories or adopted from published methods, all clinical microbiology laboratories need to be able to understand, critically evaluate, perform and interpret these tests according to rigorous and clinically appropriate standards and international guidelines. The cost and effort of development and evaluation of in-house tests is considerable and many laboratories do not have the resources to do so. This compendium is a vehicle to improve and maintain the clinical relevance and high quality of diagnostic PCR. It is a unique collection of; guidelines for PCR laboratory set up and quality control, test selection criteria, methods and detailed step by step protocols for a diagnostic assays in the field of molecular microbiology. The structure of the book provides the PCR fundamentals and describes the clinical aspects and diagnosis of infectious disease. This is followed by protocols divided into; bacteria, virus, fungi and parasites, and susceptibility screens. The inclusion of medical criteria and interpretation adds value to the compendium and benefits clinicians, scientists, researchers and students of clinical diagnostic microbiology
Recent and forecasted advances in microbiology, molecular biology, and analytical chemistry have made it timely to reassess the current paradigm of relying predominantly or exclusively on traditional bacterial indicators for all types of waterborne pathogens. Nonetheless, indicator approaches will still be required for the foreseeable future because it is not practical or feasible to monitor for the complete spectrum of microorganisms that may occur in water, and many known pathogens are difficult to detect directly and reliably in water samples. This comprehensive report recommends the development and use of a "tool box" approach by the U.S Environmental Protection Agency and others for assessing microbial water quality in which available indicator organisms (and/or pathogens in some cases) and detection method(s) are matched to the requirements of a particular application. The report further recommends the use of a phased, three-level monitoring framework to support the selection of indicators and indicator approaches.Â
This text on viable but non-culturable organisms provides information on topics including: morphological changes; the role of membranes; genetics and genetic regulation; molecular methods for detection; as well as survival dominancy and related phenomena. The main purpose of the text is to elucidate the phenomenon and to distinguish it from other seemingly related but different phenomena such as spore formation, dormancy, starvation, and injury. It covers a cross section of morphology, metabolism, genetics, ecology and epidemiology.
Presents a state-of-the-art review of the current technology and applications being utilized to identify sources of fecal contamination in waterways. - Serves as a useful reference for researchers in the food industry, especially scientists investigating etiological agents responsible for food contamination. - Provides background information on MST methods and the assumptions and limitations associated with their use. - Covers a broad range of topics related to MST, including environmental monitoring, public health and national security, population biology, and microbial ecology. - Offers valuable insights into future research directions and technology developments.
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
This second volume focuses on PCR methods and PCR application specificities to the biotechnology and bioengineering field. New and updated chapters detail real-time PCR protocols, synthetic biology applications, pathogen detection, microfluidics, digital, multiplex detection recent advances. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, PCR: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
Public Health Microbiology: Methods and Protocols is focused on microorganisms that can present a hazard to human health in the course of everyday life. There are chapters dealing with organisms that are directly pathogenic to humans, including bacteria, viruses, and fungi; on organisms that produce toxins during growth in their natural habitats; on the use of bacteriocins produced by such organisms as lactobacilli and bifidobacteria; as well as several chapters on hazard analysis, the use of disinfectants, microbiological analysis of cosmetics, and microbiological tests for sanitation equipment in food factories. Additional chapters look at the use of animals (mice) in the study of the various characteristics of milk and their relationships with lactic acid bacteria in particular. Other chapters focus on special methods for determining particular components of milk. In particular, in Parts I and II, on bacterial and viral pathogens, special attention is given to methods for PCR detection of genes with resistance to tetracycline, as well as to Salmonella enterica; for identification and typing of Campylobacter coli; for detection of the abundance of enteric viruses, hepatitis A virus, and rotaviruses in sewage, and of bacteriophages infecting the O157:H7 strain of Escherichia coli. Part III offers methods for computerized analysis and typing of fungal isolates, for isolation and enumeration of fungi in foods, and for the determination of aflatoxin and zearalenone.
As more original molecular protocols and subsequent modifications are described in the literature, it has become difficult for those not directly involved in the development of these protocols to know which are most appropriate to adopt for accurate identification of bacterial pathogens. Molecular Detection of Human Bacterial Pathogens addresses th