Dna Methods In Clinical Microbiology

DNA Methods in Clinical Microbiology describes the novel DNA-based technology now used in the diagnosis and management of infectious diseases. It is a concise, yet readable, overview written primarily for clinicians, clinical microbiologists, medical students and undergraduates in medical and veterinary microbiology. The book has two primary aims. First, to explain the principles of these methods at the `molecular' level. Second, to provide a clinical perspective by reporting results from actual DNA-based investigations on a range of specimens. Those approaching DNA methods for the first time are assisted by a brief résumé of the relevant features of nucleic acids (Chapter 2): this information is essential for an understanding of later chapters. Subsequent text covers detection, characterization and quantification of pathogens by a variety of methods - e.g., target amplification (PCR, LCR, NASBA, TMA and SDA), signal amplification (bDNA) and probe-based techniques; the chapter on typing describes nearly twenty named molecular methods, including spoligotyping and MLST. All chapters include an adequate range of current reference from which, if required, detailed protocols can be obtained. The diagrams are clear, and readers are assisted by a detailed index.

The accurate identification and typing of microbes is essential for workers active in all fields of microbiology. Many examples of modern molecular methods have been concealed in scientific and medical literature but this introductory text considers the possible applications of such methods and compares their advantages and disadvantages.

The papers published herein comprise the presentations given at the eighteenth of an annual series of clinical symposia arranged under the auspices of the Eastern Pennsylvania Branch of the American Society for Microbiology. This symposium allowed approximately 200 persons to gather and exchange ideas on the rapid laboratory diagnosis of infectious diseases. The institution of the Diagnosis Related Group (DRG) method for reimbursement by both government agencies and private insurance carriers has provided a financial aspect to the established clinical reasons for rapid laboratory diagnosis. Now the health of the institution, as well as the patient, is dependent on a timely diagnosis and, hopefully, cure. Accordingly, the goal of this symposium was to present the latest developments in "same-day microbiology". In the face of stable or diminishing resources, the laboratory director is presented with many choices. Do nucleic acid probes, non instrumental ELISA techniques, or time-resolved fluorometry have a place in his or her laboratory? Should the laboratory test for newly described human pathogens such as human immunodeficiency virus or human papilloma virus? Can rapid techniques supplant conventional methods? Or are they merely adjunctive? This symposium attempted to assist in the formulation of informed decisions. Bruce Kleger Donald Jungkind Eileen Rinks Linda A. Miller vii ACKNOWLEDGEMENTS We would like to thank the Eastern Pennsylvania Branch of t~e American Society for Microbiology for sponsoring this symposium and for making this publication possible. We especially thank the Symposium Committee for their diligent work in organizing an informative and successful symposium.

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

In the United States, hospitals annually report over 5 million cases of infectious-disease-related illnesses: clinical microbiology laboratories in these hospitals are engaged in detecting and identifying the pathogenic microorganisms in clinical specimens collected from these patients with suspected infections. Clearly, the timely and accurate detection/identification of these microbial pathogens is critical for patient treatment decisions and outcomes for millions of patients each year. Despite an appreciation that the outcome of an infectious-disease-related illness is directly related to the time required to detect and identify a microbial pathogen, clinical microbiology laboratories in the United States as well as worldwide have long been hampered by traditional culture-based assays, which may require prolonged incubation time for slowly growing microorganisms such as Mycobacterium tuberculosis. Moreover, traditional culture-based assays often require multiple steps with additional time needed for discernment of species and/or detection of antimicrobial resistance. Finally, these traditional, slow multistep culture-based assays are labor-intensive and required skilled clinical microbiologists at the bench. Over the past several decades, advanced molecular techniques in diagnostic microbiology quietly have been revolutionizing the practice of clinical microbiology in the hospital setting. Indeed, molecular diagnostic testing in general and nucleic-acid-based amplification methods in particular have been heralded as diagnostic tools for the new millennium. There is no question that the development of rapid molecular techniques for nucleic acid amplification/characterization combined with automation and user-friendly software has greatly broadened the diagnostic capabilities of the clinical microbiology laboratory. These technical advances in molecular microbiology over the first decade of the 21st Century have profoundly influenced the physical structure of clinical microbiology laboratories as well as their staffing patterns, workflow, and turnaround time. These molecular microbiology advances have also resulted in the need for a revised and updated second edition of Advanced Techniques in Diagnostic Microbiology. This second edition again provides an updated and comprehensive description of the ongoing evolution of molecular methods for the diagnosis of infectious diseases. In addition, many new chapters have been added, including a chapter on the clinical interpretation and relevance of advanced technique results. The second edition, like the first edition, includes both a “techniques” section describing the latest molecular techniques and an “applications” section describing how these advanced molecular techniques are being used in the clinical setting. Finally, the second edition, like the first edition, utilizes a diverse team of authors who have compiled chapters that provide the reader with comprehensive and useable information on advanced molecular microbiology techniques.

The Manual of Commercial Methods in Clinical Microbiology 2nd Edition, International Edition reviews in detail the current state of the art in each of the disciplines of clinical microbiology, and reviews the sensitivities, specificities and predictive values, and subsequently the effectiveness, of commercially available methods – both manual and automated. This text allows the user to easily summarize the available methods in any particular field, or for a specific pathogen – for example, what to use for an Influenza test, a Legionella test, or what instrument to use for identification or for an antibiotic susceptibility test. The Manual of Commercial Methods in Clinical Microbiology, 2nd Edition, International Edition presents a wealth of relevant information to clinical pathologists, directors and supervisors of clinical microbiology, infectious disease physicians, point-of-care laboratories, professionals using industrial applications of diagnostic microbiology and other healthcare providers. The content will allow professionals to analyze all commercially available methods to determine which works best in their particular laboratory, hospital, clinic, or setting. Updated to appeal to an international audience, The Manual of Commercial Methods in Clinical Microbiology, 2nd Edition, International Edition is an invaluable reference to those in the health science and medical fields.

In recent years, advanced molecular techniques in diagnostic microbiology have been revolutionizing the practice of clinical microbiology in the hospital setting. Molecular diagnostic testing in general and nucleic acid-based amplification methods in particular have been heralded as diagnostic tools for the new millennium. This third edition covers not only the most recent updates and advances, but details newly invented omic techniques, such as next generation sequencing. It is divided into two distinct volumes, with Volume 1 describing the techniques, and Volume 2 addressing their applications in the field. In addition, both volumes focus more so on the clinical relevance of the test results generated by these techniques than previous editions.

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.