疾病及处理

One of the continuing objectives of molecular biology research is to characterize the functronal domains of proteins. Many proteins contam domains capable of binding specific ligands, such as cofactors, substrates, and domains of other proteins, that form the basis for interactions t ...

Most attempts to identify and isolate a novel cDNA result in the acquisition of clones that represent only a part of the mRNA’s complete sequence. The approach described here to clone the missing sequence (cDNA ends) employs polymerase chain reaction (PCR). Since the initial reports of rapid amp ...

Control of gene transcription, the process in which a gene’s DNA sequence serves as a template for mRNA synthesis, plays a critical role in the multistep process that regulates gene expression. Gene transcrtption levels within a cell change in response to a wide variety of signals that occur during ...

Recombinant proteins can be detected and characterized via several immunodetection schemes. Native proteins are typically detected by applying them to a membrane manually or using a vacuum manifold in a dot-or slot-blot format. This approach is suited to screening a number of samples for t ...

Enzyme-linked immunosorbent assays (ELISAs) can be designed to detect either antigens or antibodies. Nearly all ELISA formats require the separation of reactants from the products of the immunoassay. The product of the assay is an immune complex consisting of target ligand, the analyte, a ...

The arm of this chapter is to provide a thorough yet terse treatment of Western blotting with multiple-channel immunodetection. Principles and practical aspects of transverse electrophoresrs will be presented, and the advantages and practice of multiple-channel antibody-bas ...

Two basic methods can be used for the identification of specific antigens by their corresponding antibodies: immunoprecipitation and immunoblotting (often referred to as Western blotting) (1,2). Each has its advantages. Immunoprecipitation is likely to permit the detection of bo ...

The use of recombinant antigens and chemically synthesized peptides are the new approaches for the construction of reliable and sensrtive diagnostic assays. Moreover, in the field of virology, the use of recombinant antigens eliminates the need to handle highly hazardous material in t ...

Human monoclonal antibodies (MAbs) are more suitable than MAbs of animal origin for clinical applications because of lower hypersensitivity reactions, less formation of circulating immune complexes and lower anti-immunoglobulin responses The classical production of hum ...

Despite the multitude of different parameters currently measured in the clinical laboratory, only a minor part of these are measured by means of biosensor-based methods. Within the group of biochemical sensors, enzyme or metabolic sensors as integrated devices in clinical analyzers a ...

This chapter demonstrates a preliminary experimental approach to detect antiviral antibodies by means of a quartz crystal immunosensor. As an example of the large numbers of immunoassays currently applied in the clinical laboratory, the screening for human immunodeficiency vir ...

One of the most important classes of reagents for clinical diagnosis is antibodies, either in polyclonal preparations, as monoclonal antibodies (MAbs), or as customized reagents prepared by genetic engineering. The enormous range of antibodies produced by hybridoma and recombin ...

Rapid growth in the field of antibody engineering occurred after it was shown that functional antibody fragments could be secreted into the periplasmic space and even into the medium of Escherichia coli by fusing a bacterial signal peptide to the antibody’s N-terminus (1,2). These findings a ...

For cloning and expressing the antigen-binding variable (Fv) portion of an antibody in Escherzchia coli, vectors have been constructed that combine the two variable regions (VH and VL) with a peptide linker (1–3). The genetic information for VH and VL is generally amplified from hybridoma cel ...

A method is described to generate dendritic cells (DCs) from human peripheral blood mononuclear cells (PBMCs). The procedure involves two major steps: (1) preparation of monocytes from human PBMCs and (2) in vitro differentiation of the monocytes into DCs by growth factors and cytokines. Ce ...

Both innate resistance and acquired cell-mediated immunity are involved in an anti-Candida response. Essential components of both the arms of the immune defense against infections by Candida spp. include phagocytic cells, i.e., polymorphonuclear neutrophils (PMNs) and mononuc ...

The measurement of antibodies in the external secretions that bathe mucosal surfaces is important in understanding the host response to the opportunistic pathogen, Candida albicans and its determinants of pathogenesis at these sites. The principal immunoglobulin isotype in muc ...

It is sometimes necessary to assess the genetic relatedness of isolates to identify the origin of an infection. In addition, evidence is accumulating that drug resistance can be associated with strains from a particular clade and that strains can exhibit anatomical specificity. It may, the ...

Intraspecific differentiation of pathogenic microorganisms is a major need in epidemiological studies concerning the source and spread of infections. This requirement is paramount for those etiologic agents of infectious diseases, which are mainly grouped into one species wi ...

A general procedure is described for the analysis of gene expression of Candida albicans cultured in a mouse infection model. This technique involves first infecting mice with Candida and subsequently harvesting blood and other tissue at specific time points during infection. The tiss ...