The polymerase chain reaction (PCR) has become an increasingly important tool in microbial diagnosis in recent years, mainly because of its rapidity, high sensitivity, and specificity. In a number of instances, parti- larly with noncultivatable and slow-growing microorganisms, reliable iden- fication and differentiation became possible only after the introduction of PCR-based methods. Because the number of published procedures is large and steadily rising, there is a need for critical evaluation, comparison of performance, and even- ally also standardization of methods to enable laboratory diagnosticians to select the optimal methodology. PCR Detection of Microbial Pathogens is designed to provide an ov- view of the possibilities and problems connected with the use of PCR-based identification and detection of important bacterial and other microbial pat- gens, among them several zoonotic agents. It is intended to provide micro- ologists and biochemists the opportunity to extend their knowledge of state-of-the-art detection procedures, as well as pre-PCR sample processing and various general aspects of PCR. The chapters of this volume have been written by investigators well conversant with molecular detection and iden- fication of microorganisms. They have contributed their extensive practical experience to the protocols, not only concerning PCR methodology itself, but also the highly important area of pre-amplification sample processing.
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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.
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