Reischl Molecular Diagnosis of Infectious Diseases

Erscheinungsjahr 2008
ISBN: 978-1-59259-597-6
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 13, 629 Seiten, eBook

Reihe: Methods in Molecular Medicine

In most societies the medical field is undergoing a dramatic reorien- tion. Fundamentally new technologies in diagnosis and therapy, as well as the extension of life expectations, have increased health costs to an extent that now nears the limits of acceptability. One consequence has been a reevaluation of the need for and duration of each individual hospitalization. For diagnostic laboratories, results must now be obtained rapidly in order to include them in therapeutic decisions. Furth- more, therapeutic approaches to the control of infectious agents, limited by the number of therapeutic compounds available and by the growing perce- age of multiresistant agents, now require improvement in the precision of diagnostic approaches. The necessity for maximal reduction of the risk of infection by the development of appropriate pharmaceutical products demands even higher levels of sensitivity in diagnostic tests. Today's highly purified antigens---often from recombinantly generated antigens or peptides, monoclonal or recombinant antibodies, and more sen- tive signal-production and signal-detection systems--have greatly advanced immunologically based detection methodology. However, this approach has biological limits that further enhancement of sensitivity cannot breach. Fortunately nucleic acids have now been quite successfully added to the sp- trum of diagnostic targets. Recombinant-produced antigens and the integration of microelectronics and microfluidics offer new perspectives and possibilities.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Reischl, Udo

Inhaltsverzeichnis

Recombinant Proteins and Peptide.- Identification of Novel Diagnostic Markers by Differential Display.- Immunoprecipitation.- RT-PCR Methods and Applications.- Quantitative RT-PCR.- Cloning cDNA Ends Using RACE.- A Method for the Rapid Identification of Epitopes and Other Functional Peptide Domains.- Mapping of Linear B-Cell Epitopes on Viral Polypeptides by Multiple Peptide Synthesis and Fine Tuning Sensitivity and Specificity of the Identified Peptide Antigens for Application in Virus Diagnosis.- Amplification of Genomic DNA by PCR.- PCR-Based Cloning and Subsequent Expression of Antigenic Proteins in Escherichia coli.- Application of TEV Protease in Protein Production.- Stable Transfected Drosophila Schneider-2 Cells as a Novel Tool to Produce Recombinant Antigens for Diagnostic, Therapeutic, and Preventive Purposes.- Use of Recombinant Antigens as Diagnostic Reagents in Enzyme-Linked Immunosorbent Assays.- Generation of Recombinant Baculovirus DNA in E.coli Using a Baculovirus Shuttle Vector.- The SFV Gene Expression System.- Expression and Purification of Recombinant Proteins Using the pET System.- Procedures for the Analysis and Purification of His-Tagged Proteins.- Detection and Immobilization of Proteins Containing the 6xHis Tag.- The Use of Recombinant Fusion Proteases in the Affinity Purification of Recombinant Proteins.- Purification and Immunological Characterization of Recombinant Antigens Expressed in the Form of Insoluble Aggregates (Inclusion Bodies).- Purification of Recombinant Proteins Having High Isoelectric Points.- Immunoblotting.- Diagnostic Application of a Multiple-Channel Immunoblot Apparatus.- Immobilization of lmmunoreactants on Solid Phases for ELlSAs.- Nonradioactive Labeling and lmmunodetection of Recombinant Proteins.- Flow Cytometry.- Basic Problems of Serological Laboratory Diagnosis.- Molecular Diagnostics Resources on the Internet.- Antibodies.- Monoclonal Antibody-Based Immunoassays.- Biosensor-Based Methods in Clinical Diagnosis.- The Quartz Crystal Microbalance (QCM) as an Immunosensor.- Biomolecular Interaction Analysis.- Recent Developments in Antibody Engineering.- Cloning and Expression of Single-Chain Fragments (scFv) from Mouse and Rat Hybridomas.- Production of a Human Antibody Library in the Phage-Display Vector pSEX81.- Screening of Phage-Displayed Antibody Libraries.- Purification, Characterization, and Biotinylation of Single-Chain Antibodies.