Immunoassays in Coagulation Testing

ISBN-13: 9781461572275 / Angielski / Miękka / 2012 / 173 str.

Z. Parvez

Opis

Rapid progress in analytical methods, within the past few decades, has led to the widespread applications of newer immunological and radioimmunoassay techniques to the diagnosis and treatment of hemorrhagic and thrombotic disorders. Major advances were made to meet the multiple challenges of improving precision, accuracy, and availability of various measurements. These advances have been paralleled by the discoveries of a close relationship between biological activities and the absolute concentration of proteins that were measured by immunological techniques. This, in turn, assured the significance and usefulness of immunological methods in the management of patients. Numerous variants of immunological tests now are available, which allow us to both determine with precision minute quantities of antigenic proteins in serum and other biological fluids and differentiate the native protein from its genetically altered or degraded forms. Methods also have been designed to immunologically evaluate some serine proteases that are in complex with proteolytic inhibitors. Due to rapid progress in this field, different laboratories unavoidably become experts in one or the other approach. In the welter of possible choices, the non-expert usually is left to follow either the most recent but as yet to be confirmed method or his own anecdotal experience. This manual not only brought together various methods in current use, but it also set certain standardized criteria for the assessment of various deficiencies and abnormalities in hemostasis.

Spis treści

1. Basic Concepts in Coagulation.- Hemostasis.- Vasoconstriction.- Prostaglandins and Platelet Aggregation.- Coagulation Cascade.- Fibrinolysis.- Serine Protease Inhibitors.- 2. Pathophysiology of Coagulation Disorders.- Hemophilia-A.- Hemophilia-B (Christmas Disease).- von Willebrand’s Disease (vWd).- Disseminated Intravascular Coagulation (DIC).- Platelet Function Defects.- Hypercoagulability.- Laboratory Testing.- Conclusions.- 3. Basic Concepts in Clinical Immunology.- Antigens (Immunogens).- Antigenic Determinant Sites.- Antibodies.- Antigen Binding Sites.- Structure.- Antibody Specificity.- Monoclonal Antibodies.- 4. Review of Immunologic Techniques.- Radial Immunodiffusion (RID).- Electroimmunodiffusion (EID).- Crossed Immunoelectrophoresis (CIEP).- Turbidimetric Immunoassay (TIA).- Nephelometric Immunoassay (NIA).- Radioimmunoassay (RIA).- Immunoradiometric Assay (IRMA).- Fluoroimmunoassay (FIA).- Enzyme-linked Immunosorbent Assay (ELISA).- Thin Layer Immunoassay (TLIA).- Particle Counting Immunoassay (PACIA).- Latex Agglutination Test.- Clinical Considerations.- Selected Bibliography.- 5. Procedure for Immunoassays.- Proteins of the Coagulation System.- Fibrinogen.- Fibrinopeptide A.- Factor VIII: RAg.- Prothrombin.- Factor XII (Hageman Factor).- Protein C.- Fibronectin.- Proteins of the Fibrinolytic System.- B?15–42 Peptide.- Fibrinogen Degradation Products (FDP).- Plasminogen.- Serine Protease Inhibitors.- ?1-Antitrypsin (?1 AT).- Antithrombin-III (AT-III).- ?2-Macroglobulin (?2-M).- ?2-Antiplasmin (?2-AP).- Platelet Specific Proteins.- Platelet Factor 4 (PF4).- Beta Thromboglobulin (?-TG).- Thromboxane B2 (TxB2).- 6. Immunotechniques in Coagulation Research.- Double Immunodiffusion.- Crossed Immunoelectrophoresis.- Antibody Neutralization Assay.- Radioimmunoassay.- Immunoadsorption.- Conclusions.- 7. Quality Control.- Immunologic Reagents.- Standard Preparation.- Internal Standard.- Units of Concentration.- Procedure.- Quality Control Charts.- 8. Future Outlook.- Conclusions.- 9. Appendix A. Techniques in Antibody Production.- Antigen Purification.- Isolation of AT-III.- Isolation of Plasminogen.- Protein Determination.- Immunization.- Preparation of Antiserum.- FITC-labeling of Antibodies.- 10. Appendix B. Blood Collecting Techniques.- Double Syringe Method.- Vacutainer Method.- 11. Appendix C. Buffers and Stains.- Phosphate Buffer.- Veronal, Barbital, or Barbitone Buffer.- Veronal Acetate Buffer pH 5.6–8.3.- Phosphate Buffered Saline (PBS).- 0.05 M Tris Buffer pH 7.2–8.6.- Stains.- Coomassie Blue.- Amido Black.- Azocarmine B.- Nigrosin.- Sudan Black B.- Destaining Solution.- Appendix D. Normal Ranges of Coagulation Proteins in Plasma.- Appendix E. Manufacturers of Immunologic Reagents and Equipment.- North American Region.- European Region.- Appendix F. Glossary of Terms.- References.