Hemostasis and Thrombosis Protocols

ISBN-13: 9781489943132 / Angielski / Miękka / 2013 / 368 str.

David J. Perry; K. John Pasi

Opis

Laboratory studies in hemostasis have traditionally focused on abn- malities of platelet function or the quantitative and qualitative disorders that affect the proteins involved in blood coagulation. However, over the last 10 years there has been an explosion in our understanding of the molecular bases that underlie many of the inherited and acquired disorders of hemostasis. Many of these disorders are now routinely diagnosed and assessed by methods that involve genotypic analysis. Indeed in the late 1990s the distinction between molecular methods for research and for routine diagnosis is becoming incre- ingly blurred. The techniques and approaches that are used in hemostasis are manifold and published in isolation in a variety of publications. The aim, therefore, of this volume Hemostasis and Thrombosis Protocols is to pull together, into a single volume, the variety of techniques that are frequently used in the field of hemostasis. We have targeted this volume at laboratories who wish to move into the field of molecular hemostasis or who may already have some expe- ence in this area but wish to develop new areas of research and diagnosis. The chapters are wide-ranging and hopefully provide a broad overview of the d- fering applications in which these standard techniques can be used. Though the articles may appear relatively specific, the techniques contained within them are applicable to the study of many different disorders and we hope that they provide a series of ideas and concepts well-suited to problem solving.

Recenzje

"...The book by Perry and Pasi confirms ...that with good recipes and kits the DNA analysis of disorders of thrombosis and hemostasis is indeed feasible for many diagnostic laboratories. The book describes in great detail a variety of laboratory protocols aimed primarily at analyzing inherited disorders of the hemostatic system. These protocols vary from simple procedures to detect factor V Leiden and prothrombin 20210 to elaborate techniques for finding mutations in disorders like hemophilia A or protein S deficiency. In addition ...this book is a useful book for those attempting to introduce or maintain DNA analysis in their laboratory...laboratory technicians may find many useful suggestions to improve their work."-Haemostasis

"Each cheater is presented in a recipe format so that the methods can e reproduced. All chapters subdivided into sections for easy reading: introduction, material, methods, and notes. ...The chapters on mutation detection in clotting factor deficiencies will be useful to hemostasis laboratories that are interested in carrier detection, prenatal diagnosis, or research. The chapters on von Willbrand factor multimer analysis, factor V Leiden mutation, an prothrombin G20210A mutation will be useful to hemostasis laboratories that want to set up these important diagnostic tests...This manual does well at meeting its goal of providing a broad overview of techniques used in the analysis of hemostatic disorders and providing strategies for future problem solving. It will be a great utility to hemostasis laboratories that wish to advance their research or diagnostic capabilities."-Modern Pathology

"very valuable for the laboratory staff in charge of gene analysis of known genetic defects in known diseases of hemostasis and thrombosis." - Bloodline

Spis treści

Part I. Introduction. Hemostasis: Components and Processes K. John Pasi Part II. Basic Techniques. Isolation of DNA and RNA David J. Perry Amplification of DNA and RNA by PCR David J. Perry Direct Sequencing of PCR Products David J. Perry Solid-Phase Sequencing of Biotinylated PCR Products with Streptavidin-Coated Magnetic Beads David J. Perry Automated DNA Sequencing Helen L. Devereux Detection of DNA by Silver Staining David J. Perry and Flora Peyvandi Promoter Studies in Hemostasis Peter R. Winship and Jonathan R. K. Spray Part III. Methods of Mutational Analysis. Detection of Mutations and Polymorphisms in Clotting Factors by Denaturing Gradient Gel Electrophoresis Rainer Schwaab and Winfried Schmidt Screening for Mutations in DNA by Single-Stranded Conformation Polymorphism (SSCP) Analysis David J. Perry Screening for DNA Heteroduplexes in the Factor VII Gene Using Ethylene Glycol Gel Electrophoresis of Solvent-Treated 32P-Labeled PCR Products Peter M. Baker Detection of Mutations Causing Hemophilia A Using an In Vitro Coupled Transcription and Translation System Chike Ononye and P. Vincent Jenkins Screening for Mutations in the Human Antithrombin Gene by Hydrolink D-5000™ and MDE™ Gel Electrophoresis David J. Perry Part IV. Methods for Analyzing Inherited/Acquired Disorders of Hemostasis Detection of Mutations In Hemophilia A Patients by Chemical Cleavage of Mismatch Method Naushin H. Waseem, Richard Bagnall, Peter M. Green, and Francesco Giannelli Inversion Mutation Analysis in Hemophilia A by Restriction Enzyme Analysis and Southern Blotting Chike Ononye and P. Vincent Hemophilia B Mutational Analysis Peter M. Green Screening for Candidate Mutations Causingvon Willebrand's Disease (vWD) P. Vincent Jenkins Use of Intron 40 VNTR I in vWD Gene Tracking Mohammed S. Enayat and Gurcharan J. Surdhar Multimeric Analysis of von Willebrand Factor (vWF) Mohammed S. Enayat Identification of Mutations in the Human Factor VII Gene Peter M. Baker Molecular Analysis in Factor XI Deficiency Karen M. Johnson and John H. McVey Mutational Analysis in Antithrombin Deficiency David J. Perry Ectopic Transcript Analysis in Human Antithrombin Deficiency David J. Perry Mutational Analysis of the Human Protein C Gene Roger Luddington Analysis of the Protein S Gene in Protein S Deficiency Núria Sala and Yolanda Espinosa-Parrilla Screening for the G to A Transition at Position 20210 in the 3'-Untranslated Region (UTR) of the Prothrombin Gene Karen P. Brown Screening for the Factor V Leiden Mutation Karen P. Brown Multiplex PCR for Detection of the Prothrombin 3' UTR (C20210A) Polymorphism and the Factor V Leiden Mutation Gillian Mellars, P. Vincent Jenkins, and David J. Perry Isoelectric Focusing and Immunodetection of Plasma Antithrombin Martina Daly Characterization of Heparin Binding Variants of Antithrombin by Crossed Immunoelectrophoresis in the Presence of Heparin Martina Daly The Determination of Amino Acid Sequence Abnormalities in Proteins by HPLC Peptide Analysis David Williamson Part V. Platelet and Megakaryocyte Analysis Molecular Biological Identification and Characterization of Inherited Platelet Receptor Disorders Ramesh B. Basani, Mark Richberg, and Mortimer Poncz In Vitro Expansion of Megakaryocytes from Peripheral Blood Hematopoietic Progenitors Michael A. Thornton and Mortimer Poncz Molecular Biology Studies with Primary Megakaryocytes Yaping

Blurb

In Hemostasis and Thrombosis Protocols, leading experimentalists describe in detail their proven, cutting-edge methods for research on hemostasis and thrombosis, as well as their diagnostics. The techniques range from widely used basic assays to methods that are specialized for mutational analysis and specific disorders. Readily reproducible, these powerful methods can be used to screen for such inherited disorders as hemophilia A and B, von Willebrands Disease, Factor XI and antithrombin deficiency, protein S deficiency, factor V Leiden mutation, and other problems. The protocols reflect the many major advances that have occurred in our understanding of the molecular genetics of normal hemostasis and it variants, as well as of the coagulation cascade itself.

Suitable for both diagnostic and research laboratories, Hemostasis and Thrombosis Protocols enables experienced and novice investigators alike readily to master the complexities of research on molecular hemostasis and thrombosis, as well as work toward the development of productive new molecular diagnostics.