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Preceded by: Clinical applications of PCR / edited by Y.M. Dennis Lo, Rossa W.K. Chiu, K.C. Allen Chan. c2006.
The polymerase chain reaction (PCR) is one of the most important molecular biological methods ever devised, with numerous applications to cli- cal molecular medicine. Since its description in 1985, PCR has undergone tremendous improvements, and many variations on the basic PCR theme have been published. With such a large volume of PCR-related literature, a clinical scientist wishing to use the technique will have a difficult task loc- ing the relevant information to implement it effectively. There is thus clearly a need for an up-to-date volume with detailed protocols to facilitate the setting up of those techniques most relevant to clinical applications. Unlike some other books on this topic, Clinical Applications of PCR includes only methods that are of direct relevance in clinical settings. The book is organized in three parts: an introductory section, a section on general methodology, and a final section with specific clinical applications. The first section covers the basic principles of PCR and is most useful to those new to molecular diagnosis. The next chapter includes useful tips for setting up a PCR laboratory. Section 2 then outlines some of the most commonly used PCR-based techniques in molecular diagnosis. Section 3 includes carefully chosen examples that represent typical applications of PCR in diverse clinical fields, encompassing hematology, oncology, genetics, and microbiology.
Basic Science Methods for Clinical Researchers addresses the specific challenges faced by clinicians without a conventional science background. The aim of the book is to introduce the reader to core experimental methods commonly used to answer questions in basic science research and to outline their relative strengths and limitations in generating conclusive data. This book will be a vital companion for clinicians undertaking laboratory-based science. It will support clinicians in the pursuit of their academic interests and in making an original contribution to their chosen field. In doing so, it will facilitate the development of tomorrow’s clinician scientists and future leaders in discovery science. Serves as a helpful guide for clinical researchers who lack a conventional science background Organized around research themes pertaining to key biological molecules, from genes, to proteins, cells, and model organisms Features protocols, techniques for troubleshooting common problems, and an explanation of the advantages and limitations of a technique in generating conclusive data Appendices provide resources for practical research methodology, including legal frameworks for using stem cells and animals in the laboratory, ethical considerations, and good laboratory practice (GLP)
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
Kary Mullis was awarded a Nobel Prize for inventing the PCR technique more than a decade ago in 1993. Since its "discovery", multiple adaptations and variations of the standard PCR technique have been described. This publication aims to provide the reader with a guide to the standard PCR technique and its many available variants, with particular emphasis being placed on the role of these PCR techniques in the clinical diagnostic laboratory (the central theme of this book).
PCR is the most powerful technique currently used in molecular biology. It enables the scientist to quickly replicate DNA and RNA on the benchtop. From its discovery in the early 80's, PCR has blossomed into a method that enables everything from ready mutation of DNA/RNA to speedy analysis of tens of thousands of nucleotide sequences daily. PCR Applications examines the latest developments in this field. It is the third book in the series, building on the previous publications PCR Protocols and PCR Strategies. The manual discusses techniques that focus on gene discovery, genomics, and DNA array technology, which are contributing factors to the now-occurring bioinformatics boom. Key Features * Focuses on gene discovery, genomics, and DNA array technology * Covers quantitative PCR techniques, including the use of standards and kinetic analysis includes statistical refinement of primer design parameters * Ilustrates techniques used in microscopic tissue samples, such as single cell PCR, whole cell PCR, laser capture microdissection, and in situ PCR Entries provide information on: * Nomenclature * Expression * Sequence analysis * Structure and function * Electrophysiology * Parmacology * Information retrieval
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.
James D. Watson When, in late March of 1953, Francis Crick and I came to write the first Nature paper describing the double helical structure of the DNA molecule, Francis had wanted to include a lengthy discussion of the genetic implications of a molecule whose struc ture we had divined from a minimum of experimental data and on theoretical argu ments based on physical principles. But I felt that this might be tempting fate, given that we had not yet seen the detailed evidence from King's College. Nevertheless, we reached a compromise and decided to include a sentence that pointed to the biological significance of the molecule's key feature-the complementary pairing of the bases. "It has not escaped our notice," Francis wrote, "that the specific pairing that we have postulated immediately suggests a possible copying mechanism for the genetic material." By May, when we were writing the second Nature paper, I was more confident that the proposed structure was at the very least substantially correct, so that this second paper contains a discussion of molecular self-duplication using templates or molds. We pointed out that, as a consequence of base pairing, a DNA molecule has two chains that are complementary to each other. Each chain could then act ". . . as a template for the formation on itself of a new companion chain, so that eventually we shall have two pairs of chains, where we only had one before" and, moreover, " ...