The era of molecular pathology has arrived. From its promising beg- nings in research laboratories, the field has grown, and continues to grow, to become a vital part of the care of an ever-increasing number of patients. Because of its recent emergence from the research taboratory, many molecuIar pathology protocols we still to be found in the primary litcramre, and have not appeared in a text. MO~PCU~Q~ Padhoiogy Protocob contains la- ratory protocoIs that have been developed by many of the authors for use in clinical molecular pathology laboratories and describe in detail Row to perform these assays. This book is therefore intended for clinical laboratory use by medical technologists and pathologists. It will dso be of interest to research workers who are performing these assays. In its broadest meaning, pathology is the study of disease, and therefore it follows that any disease for which the molecular basis is understood would be suitable as a topic for inclusion in this work. When seiecting protocols, it was necessary to place limits on the number of chapters that could be feasibly presented in a single work. Those protoculs that were selected are performed more frequently, or have achieved recognition as having important diagnostic utility in contemporary practice. A decision was made to exclude inherited genetic diseases with certain exceptions, such as those diseases that are associated with thrombotic states and are part of the traditional dumain of pathology.

We are currently experiencing a fundamental shift in the way in which we approach the characterization of cancer. Never before has the make up of cancer tissues and individual cells been so exhaustively researched and char- terized. We are now capable of producing molecular “fingerprints” that ch- acterize the expression of all known and unknown genes within tumors and their surrounding tissues. More than 30,000 different genes may be measured in each patient’s tumor in a single experiment. Simultaneously, novel therapies that exploit the molecular roadmap have been developed and are now being offered to patients. These novel agents, such as Glivec, Herceptin, Iressa, and others, specifically target individual genes within tumors and can produce d- matic responses in some patients. These drugs are only the forerunners of a coming tidal wave of novel therapeutics that individually target specific m- ecules within cancer cells—more than 300 such agents are currently in phase I or II clinical trials. This is an exciting time for cancer specialists and patients alike. However, if we have learned anything from the past 50 or more years of research into cancer, it is that Lord Beaverbrook, in founding the British national health service in the 1950s, was frighteningly prescient when he defined the primary goal of health care to be “Diagnosis, Diagnosis, Diag- sis. ” Now, more than ever, it is essential that appropriate diagnostic methods and approaches are applied to the selection of patients for treatment.

Anthony Killeen, MD, PhD, offers a comprehensive yet concise introduction to molecular pathology that encompasses both applied and theoretical knowledge. Writing in a very readable style, the author reviews the basic concepts of human molecular biology, explains the principles of the most commonly used analytical methods, and discusses the molecular principles that underlie both inherited diseases and acquired genetic abnormalities that lead to cancer. Using common viral infections as examples, the book applies these molecular methods to the detection of microbial pathogens. The growing importance of pharmacogenetics and identity testing in the clinical laboratory is also highlighted.

A huge amount of fixed and paraffin-embedded tissue is stored in every hospital. This is very precious material that can be used for translational research and for diagnostics. The molecular methods employed for analysis of these tissues are similar to the usual molecular biology and proteomics methods, but reliable results can be obtained only if specific steps are followed with great care. This book provides detailed and precise guidelines for molecular analysis of archive tissues and will serve as an invaluable aid for researchers and pathologists involved in translational research and diagnostics. Clear notes and explanations are included to simplify use of the protocols for the less experienced. The authors are a group of acknowledged experts who have developed the described methods and validated them within the European project "Archive Tissues: Improving Molecular Medicine Research and Clinical Practice - IMPACTS", which has involved 21 leading institutions in 11 countries.

This is the first handbook to provide an all-in-one guide to establishing molecular biology protocols with requisite quality control. Molecular Biology and Pathology will help professionals sift through the incredible wealth of information available on molecular biology, specifically as it relates to the clinical arena of molecular pathology. This handbook provides excellent training information, and the concern of safety is discussed extensively. The handbook can serve as a primer and reference for those interested in the technical topics described, including the brief discussion of DNA banking. Quality Control (QC) suggestions are also presented.

Anthony Killeen has assembled a collection of readily reproducible molecular pathology techniques that are either frequently performed or recognized for their significant diagnostic utility. Each method is described in step-by-step detail by a leading molecular pathologist or laboratory scientist who has developed it or used it extensively. These clinical laboratory techniques can be used for the diagnosis or monitoring of cancer, hematological malignancies, infectious diseases, and selected genetic disorders. Comprehensive and path-breaking, Molecular Pathology Protocols will enable clinical laboratories to introduce new molecular pathology tests and lay the groundwork for a much-needed standardization in this rapidly developing field.

Major advancements in the field of in situ molecular pathology have occurred since publication of the first edition. In Situ Molecular Pathology and Co-expression Analyses, Second Edition, continues to teach both the molecular basis for the improvements and the actual protocols. This is the unique feature that separates it from the pack of other "cook-book" type approaches. The fields of in situ hybridization and immunohistochemistry have expanded rapidly where computer-based analyses systems have greatly expanded the power of these methods. Further, knowledge of the marked improvements in the reagents themselves since the first edition can make the difference of excellent versus misleading data. The automated platforms require that researchers and diagnostic biomedical investigators have a good understanding of the basics of in situ based tests, protocols, and biochemistry for troubleshooting in order to maximize the use of these platforms. This second edition focuses attention on straightforward protocols used to simultaneously detect two or more proteins/nucleic acids within intact tissue by doing co-expression analyses. Practicing molecular pathologists, diagnostic pathologists, laboratory directors, and toxicologists, as well as clinicians and researchers in training, will benefit from this clear presentation of protocols and theoretical framework. Data derived from in situ hybridization and immunohistochemistry. - Explains the theory and foundation of immunohistochemistry and in situ hybridization and presents easy-to-follow experimental protocols with tricks of the trade - Includes two new chapters: Recent improvements in immunohistochemistry and in situ hybridization, Quality control for immunohistochemistry and in situ hybridization: How to know if the color change is signal or background - The second edition also includes a detailed test to help one learn the basics of histologic interpretation of tissues and a separate detailed test in how to differentiate signal from background - Includes chapter-ending summaries of Key Points to Remember, bringing beginners up to speed with any seasoned veteran in the field - Thoughtfully tackles the molecular basis if IHC and ISH, along with application of that knowledge to improving the techniques is significant

This essential methods manual for immunohematologists (or hematologists and immunohematologists) provides information on genes that encode antigens on red blood cells, platelets and neutrophils. The book begins by covering general concepts in molecular biology and specific protocols such as DNA preparation, PCR-RFLP and allele-specific PCR. Information on the erythrocyte, platelet and neutrophil antigen systems and the molecular basis of polymorphisms are presented clearly in a gene facts sheet format. Database accession numbers and useful adjuncts such as Request forms, worksheets for PCR/enzyme digests also serve to benefit the user. The information is clearly presented and easily accessible and is complemented by the excellent diagrams and tabular material. This book is invaluable for both new and experienced researchers in the field and other related disciplines. - Essential for hematologists and those involoved in tissue typing and the study of human genetic polymorphisms - Presents clearly and concisely the information on a particular variant and the technique used to detect it - Organized by antigen and provides sequences of polymorphisms and primers - Details the general concepts and critical information on genes, their products, and sources of relevant nucleic acids - Includes protocols that allow investigators to set up assays with minimal effort (protocols include primers, reagents, reaction conditions, sizes of amplified products, restriction fragment digests, and the relevant safety information) - Provides information that helps interpret results in clinical settings - Contains additional sources of information (e.g., key references, web site addresses, glossary, Database accession numbers, request forms, and worksheets for PCR/enzyme digests)