Molecular Diagnosis of Infectious Diseases

Populations of the western world are now healthier and enjoying higher life expectancy than ever. They are beginning to benefit from an array of costly new therapies made possible through recent rapid advances in medical science and technology, and their demands on modern medicine are rising. Meanwhile, healthcare systems are struggling with their outdated legacy models of the m- th 20 century and are experiencing ever-increasing financial pressure from g- ernments and health insurance organizations. The equation is no longer in balance, and this predicament is forcing societies to explore new approaches to managing healthcare in the future. Since the first edition of Molecular Diagnosis of Infectious Diseases was published, we have witnessed the sequencing of the (almost) complete human genome and a shift in medical research from an emphasis on genetics to the advancement and useful application of proteomics. Bioinformatics has become the key tool for managing and analyzing the upsurge of data, and faster and more effective test methods and technologies have opened up new prospects for ind- try and academia. The tools of modern genomics and proteomics are now being utilized to specifically guide the discovery of drugs for the prevention, diagnosis, and treatment of human disease. They may also help us to find a way out of the current healthcare calamity.

Includes supplementary material: sn.pub/extras

Klappentext
In this second edition of a much-acclaimed classic laboratory manual, Molecular Diagnosis of Infectious Diseases, leading experts from academia and industry describe cutting-edge methods for the protein-based diagnosis of infectious diseases. Explaining the latest developments in genomics, proteomics, bioinformatics, biosensors, high-throughput devices, and recombinant technology, the authors demonstrate the power of these new methodologies in both established and novel procedures by applying them successfully to the identification and characterization of valuable diagnostic markers, immunomodulatory components, epitope mapping, the production and purification of recombinant antigens, and their use in diagnostic reagents in immunological assays. Basic problems of serological laboratory diagnostics and the use of immunoproteomics for discovering target proteins are also addressed. Each readily reproducible protocol is described in step-by-step detail and includes a background introduction, equipment and reagent lists, tips on troubleshooting and avoiding known pitfalls, and extensive references.
Comprehensive and up-to-date, Molecular Diagnosis of Infectious Diseases, Second Edition offers novice investigators an easy entry into the field and experienced researchers a broad array of time saving tools that will not only elucidate new principles, but also greatly facilitate the development and successful application of rapid diagnostic methods.

Inhalt
Protein-Based Identification Of Antigens.- Proteomic Approaches to Antigen Discovery.- Immunoproteomics.- Immunoprecipitation and Blotting.- DNA-Based Identification of Antigens.- Representational Difference Analysis of cDNA.- Microarray Data Analysis and Mining.- Expression Cloning.- Identification of Epitopes and Immunomodulatory Components.- Determination of Epitopes by Mass Spectrometry.- Identification of T-Cell Epitopes Using ELISpot and Peptide Pool Arrays.- Virus-like Particles.- Application of Single-Cell Cultures of Mouse Splenocytes as an Assay System to Analyze the Immunomodulatory Properties of Bacterial Components.- Expression of Recombinant Proteins.- High-Throughput Expression and Purification of 6xHis-Tagged Proteins in a 96-Well Format.- Production of Antigens in Chlamydomonas reinhardtii.- Codon-Optimized Genes that Enable Increased Heterologous Expression in Mammalian Cells and Elicit Efficient Immune Responses in Mice after Vaccination of Naked DNA.- Purification, Modification, and Renaturation of Recombinant Proteins.- Purification and Immunological Characterization of Recombinant Antigens Expressed in the Form of Insoluble Aggregates (Inclusion Bodies).- Purification of Recombinant Proteins with High Isoelectric Points.- Refolding of Inclusion Body Proteins.- Small-Molecule-Protein Conjugation Procedures.- Characterization of Recombinant Proteins.- Structural Characterization of Proteins and Peptides.- Determination of Kinetic Data Using Surface Plasmon Resonance Biosensors.- Affinity Measurements of Biological Molecules by a Quartz Crystal Microbalance (QCM) Biosensor.- Evaluation of Recombinant Proteins in Immunological Test Systems.- Solid Supports in Enzyme-Linked Immunosorbent Assay and Other Solid-Phase Immunoassays.- Design and Preparationof Recombinant Antigens as Diagnostic Reagents in Solid-Phase Immunosorbent Assays.- Basic Problems of Serological Laboratory Diagnosis.- Molecular Diagnostics Resources on the Internet.- Recombinant Receptor Molecules.- Cloning Single-Chain Antibody Fragments (scFv) from Hybridoma Cells.