PCR Detection of Microbial Pathogens Methods in Molecular Biology Hardcover

Because the number of published PCR procedures is already large and rising steadily, there is a need to critically evaluate and eventually standardize its methodology so that laboratory diagnosticians can select optimal techniques for their own work. In PCR Detection of Microbial Pathogens, 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. Described in step-by-step detail to ensure successful experimental results, 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. Each article details a validated technique that explains the principles involved, provides stepwise instructions and a list of materials, and includes notes describing problems that may arise, as well as possible alternative procedures. Authoritative and easy-to-use, PCR Detection of Microbial Pathogens offers microbiologists, veterinary diagnosticians, and food analysts powerful state-of-the-art tools designed not only to expand their knowledge of the potential of DNA-based diagnostic methods, but also to help select those techniques best suited to their specific practical needs. Specificity And Performance Of Diagnostic Pcr Assays / K. Sachse -- Pre-pcr Processing Of Samples / P. Radstrom [and Others] -- Critical Aspects Of Standardization Of Pcr / J. Hoorfar And N. Cook -- Pcr Technology And Applications To Zoonotic Food-borne Bacterial Pathogens / P.s. Lubeck And J. Hoorfar -- Detection, Identification, And Subtyping Of Actinobacillus Pleuropneumoniae / J. Frey -- Identification And Differentiation Of Brucella Abortus Field And Vaccine Strains By Bass-pcr / D.r. Ewalt And B.j. Bricker -- Isolation Of Campylobacter And Identification By Pcr / M.d. Englen, S.r. Ladely And P.j. Fedorka-cray -- Detection And Differentiation Of Chlamydiae By Nested Pcr / K. Sachse And H. Hotzel -- Detection Of Toxigenic Clostridia / M.r. Popoff -- Pcr-based Detection Of Coxiella Burnetii From Clinical Samples / M. Berri, N. Arricau-bouvery And A. Rodolakis -- Detection And Subtyping Of Shiga Toxin-producing Escherichia Coli (stec) / P. Gallien --^ Detection Of Listeria Monocytogenes Using A Pcr/dna Probe Assay / L. O'connor -- Detection Of Leptospira Interrogans / J.w. Lester And R.b. Lefebvre -- Detection Of Pathogenic Mycobacteria Of Veterinary Importance / R.a. Skuce [and Others] -- Multiplex Pcr Of Avian Pathogenic Mycoplasmas / M.i. Khan -- Detection And Differentiation Of Ruminant Mycoplasmas / H. Hotzel [and Others] -- Detection Of Mycoplasma Hyponeumoniae From Clinical Samples And Air / M. Kobisch And J. Frey -- Pcr-detection Of Hemophilus Paragallinarum, Hemophilus Somnus, Mannheimia (pasteurella) Hemolytica, Mannheimia Spp., Pasteurella Trehalosi, And Pasteurella Multocida / H. Christensen [and Others] -- Detection Of Salmonella Spp / B. Malorny And R. Helmuth -- Detection Of Toxoplasma Gondii / J.m. Wastling And J.g. Mattsson -- Pcr-derived Methods For The Identification Of Trichinella Parasites From Animal And Human Samples / E. Pozio And G. La Rosa --^ Detection Of Pathogenic Yersinia Enterocolitica By A Swab Enrichment Pcr Procedure / R. Knutsson And P. Radstrom -- Pcr Detection Of Microbial Pathogens. Introduction / K. Sachse And J. Frey. Edited By Konrad Sachse And Joachim Frey. Includes Bibliographical References And Index. 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 Presents a set of "classic" PCR-based methods for the identification and detection of animal and food microbial pathogens, including several zoonotic agents. This work covers chapters that review the specificity and performance of diagnostic PCR analysis, the pre-PCR processing of samples, and the critical aspects of standardizing PCR methods. The undisputed success of detection assays based on the polymerase chain reaction (PCR) has been largely due to its rapidity in comparison to many conventional diagnostic methods.