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Immunochemical Techniques
Time of Update: 2021-02-20
This chapter contains the practical basics of conjugation (a large field in itself), and details other immediately useful techniques that might be first desired in starting an ELISA.
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Systems in ELISA
Time of Update: 2021-02-20
Such a chapter is important because the possibilities inherent in the systems of ELISA must be understood in order to maximize their versatility in assay design.
2. Separation of bound and free reagents, which are added subsequently to the solid phase-attached substance, is by a simple washing step.
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Molecular Genotyping of Irish Rotavirus Strains
Time of Update: 2021-02-20
In developing countries, it is estimated that rotavirus is responsible for one-third of all diarrhea-associated hospitalizations and 873,000 deaths annually ( 2 , 3 ).
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Genomic DNA Digestion and Ribotyping
Time of Update: 2021-02-20
Such studies generally mvolve characterization to the genus and species level and provide an identification for the organism.
The practice of obtaining these patterns (or profiles) for epidemiologic purposes is the subject of the present chapter, although the methods can also be applied to questions regarding identification at higher taxonomic levels.
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Selection of Recombinant Baculoviruses by Visual Screening
Time of Update: 2021-02-20
The baculovirus expression system is now a widely accepted tool for the expression of recombinant proteins with many features to recommend it.
Examples of glycosylation ( 1 ), phosphorylation ( 2 ), myristilation ( 3 ), palmitoylation ( 4 ), and carboxy-methylation ( 5 ) are among the many modifications reported.
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Molecular Aspects of Disease Pathogenesis in the Transmissible Spongiform Encephalopathies
Time of Update: 2021-02-20
Over the last 20 yr, they have gone from a fascinating but relatively obscure group of diseases to one that is a major agricultural and economic problem as well as a threat to human health.
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Assay of mtDNA Polymerase from Human Tissues
Time of Update: 2021-02-20
MitochondrialDNA (mtDNA) replication is a complex process involving over 20 proteins organized along the inner mitochondrial membrane as a multienzyme complex called the mtDNA replisome, or replication factory (1 –3 ).
Figure 1 illustrates some of the protein components that participate in mitochondrial DNA replication.
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Transcriptomic Analyses of Saliva
Time of Update: 2021-02-20
In this chapter, we will overview the development of sensitive and robust microarray and multiplex quantitative reverse transcriptase- PCR assays for the discovery and validation of mRNA biomarkers in human saliva.
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Detection of Listeria monocytogenes by the Nucleic Acid Sequence-Based Amplification Technique
Time of Update: 2021-02-20
The amplification in a typical PCR process can result in a million-fold increase in the number of target sequence copies after 20 cycles ( 1 ).
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Use of Escherichia coli Mutator Cells to Mature Antibodies
Time of Update: 2021-02-20
Repeated rounds of mutation and increasingly stringent selection (Fig. 1 ) enable recovery of Abs of substantially elevated affinity for the target.
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Detection and Molecular Epidemiology of Rotavirus by RNA Gel Electrophoresis
Time of Update: 2021-02-20
The rotaviruses contain a genome of 11 segments of double-stranded RNA that can be separated into distinct bands by electrophoresis.
Electrophoretic separation of the segmented genome has gained popularity as a method not only for detection of rotavirus but also for molecular epidemiological studies (1 ,2 ).
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Monkey B virus (HS) enzyme-linked immunology analysis (ELISA)
Time of Update: 2021-02-20
Font-SIZE: < 10.5pt;">450 < FONT-SIZE: 10.5pt; FONT-FAMILY: "New Times Roman"; '>nm wavelength measurement absorbent ( OD value), through the standard curve >; B virus (HS) concentration.
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Multiplex PCR for Specific Identification of Enterohemorrhagic Escherichia coli Strains in the O157:H7 Complex
Time of Update: 2021-02-20
The “O157:H7 complex” is comprised mostly of enterohemorrhagicEscherichia coli (EHEC) strains, with serotype O157:H7 being the prototypic and predominate pathogenic strain in the complex.
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Determination of the Transforming Activities of Adenovirus Oncogenes
Time of Update: 2021-02-20
Today it is well established that cell transformation by human Ads is a multistep process involving several gene products encoded in early transcription units 1A ( E1A ) and 1B ( E1B ).
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Determination of Acyl Homoserine Lactone and Tetramic Acid Concentrations in Biological Samples
Time of Update: 2021-02-20
This process is especially prominent in the pathogenicity ofPseudomonas aeruginosa , an opportunistic human pathogen that forms sessile communities known as biofilms.
As such, a method for the detection of these QS signals may provide insights into the pathogenicity and survival ofP.
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Pneumococcal Diseases
Time of Update: 2021-02-20
The pneumococcus is now the most important bacterial pathogen causing pneumonia, meningitis, and otitis media. Traditional approaches to the diagnosis ofStreptococcus pneumoniae as the etrologic age
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Directed Evolution of Carotenoid Synthases for the Production of Unnatural Carotenoids
Time of Update: 2021-02-20
Directed evolution is a well-established strategy to confer novel catalytic functions to the enzymes.
Thanks to the relative ease of establishing color screening, carotenogenic enzymes can be rapidly evolved in the laboratory for novel functions.
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Analysis of the DNA of Parasitic Protozoa by Flow Cytometry
Time of Update: 2021-02-20
Parasitic protozoa, generally, present as highly disperse popula tions with complex life cycles.
The ability to attain and sustain this diversity and complexity resides in theDNA of the cell.
There are three methodologies that can be used to study intact cell or organellar DNA: colorimetry, microspectrofluorometry, and flow cytometry.
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Analysis of DNaseI Hypersensitive Sites in Chromatin by Cleavage in Permeabilized Cells
Time of Update: 2021-02-20
Treatment of nuclei with limited amounts of DNaseI can be used to reveal sites in chromatin that are hypersensitive (HS) to the nuclease ( 1 , 2 ).
Mapping of DNaseI HS sites and monitoring their time course of appearance (or disappearance) has been successfully used to identify many regulatory elements.
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Genome-Wide Transcriptome Analysis in Yeast Using High-Density Tiling Arrays
Time of Update: 2021-02-20
This protocol will enable researchers not only to study which genes are expressed and to what levels, but also to identify non-coding RNAs and to study the structure of transcripts including their untranslated regions, alternative start, stop, and processing sites.
