PCR Detection of Microbial Pathogens: (943 Methods in Molecular Biology)

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 Biology™ 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.

Table of Contents:
Real-time Quantitative PCR, Pathogen Detection and MIQE.- Overcoming Inhibition in Real-time Diagnostic PCR.- Quality in the Molecular Microbiology Laboratory.- Pre-Analytical Sample Treatment and DNA Extraction Protocols for the Detection of Bacterial Pathogens from Whole Blood.- Detection of Bacterial Contamination in Platelet Concentrates using Flow Cytometry and Real-time PCR Methods.- Multiplex Real-time PCR Assay for the Detection of Meticillin-Resistant Staphylococcus aureus and Panton-Valentine Leukocidin from Clinical Samples.- PCR Detection of Haemophilus influenzae from Respiratory Specimens.-  Rapid Detection of Atypical Respiratory Bacterial Pathogens by Real-time PCR.- Use of Multiple Real-time PCR Assays for the Diagnosis of Bodetella spp in Clinical Specimens.- Detection of Mycoplasma pneumoniae by Real-time PCR.- Real-Time PCR Assay for the Diagnosis of Pneumocystis jirovecii Pneumonia.- Rapid Identification of Mycobacteria and Rapid Detection of Drug Resistance in Mycobacterium tuberculosis in Cultured Isolates and in Respiratory Specimens.- Direct Detection of Mycobaterium ulcerans in Clinical Specimens and Environmental Samples.- Detection of Bartonella spp. DNA in Clinical Specimens using an Internally Controlled Real-time PCR Assay.- Direct Identification of Multiple Genital Pathogens in Urine using Multiplex PCR-based Reverse Line Blot Assays.- Diagnosis of Clostridium difficile Infection using Real-time PCR.- Detection of Pathogenic Leptospira spp. through Real-time PCR (qPCR) Targeting the LipL32 Gene.- Sensitive and Rapid Detection of Campylobacter jejuni and Campylobacter coli using Loop-mediated Isothermal Amplification.- PCR Detection of Helicobacter pylori in Clinical Samples.- Rapid Detection of the Escherichia coli Genospecies in Water by Conventional and Real-time PCR.- Multiplex Real Time PCR (MRT-PCR) for Diarrheagenic Escherichia coli.