微生物学技术

When attempting to establish libraries of immunoglobulins (Igs) from human subjects during the course of infection or other illness, a number of basic problems present themselves. First, the only source of lymphocytes that can be easily sampled is the peripheral blood in which the represen ...

The development of phage-display technology and the construction of huge libraries of antibody (Ab) fragments have provided an unlimited source of binders to virtually any antigen (Ag) (1). However, it is unlikely that the heavy (VH) and light (VL) chains of the Abs obtained from these libraries r ...

Recombinant antibody (Ab) libraries have been constructed from a wide range of B-lymphocyte sources using a number of different approaches. Sizes of the libraries that have been produced vary considerably, from small libraries of 106 up to large libraries1010. Often an Ab with the desired sp ...

Food, water, pharmaceutical, and medical microbiology ideally need methods of detecting, identifying, and quantifying microorganisms that can give results within a few hours. The many days needed for conventional enrichment, plating, and biochemical/serological methods to g ...

Many areas of analytical microbiology that deal with biological materials (e.g., medicine and the food, water, and pharmaceutical industries) need methods of detecting and identifying microorganisms that can give results in a few hours rather than the many days required for convention ...

To understand the principle of an immunoassay, one must be familiar with the antibody-antigen relationship. An antigen is a molecule that, when injected into an animal, will elicit an antibody response. Antigens may be proteins, polypeptides, or carbohydrates (glycolipids or glycopro ...

The isolation of genomic DNA from a microorganism generally comprises three stages: cultivation of the cells, disruption to release cell contents, and chemical purification of the DNA. Two widely used methods for the preparation of bacterial DNA are those described by Marmur (1) and Kirby (2), ...

This chapter outlines the basic requirements for finding and exploring sequences of interest in public databases, such as GenBank. As such, it is not aimed at experienced sequencers, for whom this will be “second nature,” but at the many clinical bacteriologists who rarely have need of DNA sequen ...

Comparative genomics is a science in its infancy. It has been driven by a huge increase in freely available genome-sequence data, and the development of computer techniques to allow whole-genome sequence analyses. Other approaches, which use hybridization as a method for comparing the gene ...

Many microbial genome sequences have been determined, and more new genome projects are ongoing. Shotgun sequencing of randomly cloned short pieces of genomic DNA can provide a simple way of determining whole genome sequences. This process requires sequencing of many fragments, compil ...

Living organisms are defined by the genes they possess. Control of expression of this gene set, both temporally and in response to the environment, determines whether an organism can survive changing conditions and can compete for the resources it needs to reproduce. Bacteria are no exceptio ...

The plethora of data now available from bacterial genome sequencing has opened a wealth of new research opportunities. Many of these have been reviewed in preceding chapters. Genomics alone, however, cannot capture a biological snapshot from an organism at a given point in time. The genome its ...

Microbiology has entered the postgenomic era and it is clear that bacterial typing should aim to be based on analysis of complete genomes. Although complete genome sequencing for epidemiological typing remains unrealistic for the present, microarrays that provide information on ge ...

Diagnostic medical bacteriology consists of two main components: identification and typing. Molecular biology has the potential to revolutionize the way in which diagnostic tests are delivered in order to optimize the care of infected patients, whether they are in hospital or in the com ...

Nucleic acid amplification technologies (NAATs) represent powerful tools in clinical microbiology, particularly in areas where traditional culture-based methods alone prove insufficient. A notable advantage is in reducing the time from taking samples to reporting result ...

This chapter describes two key strategies for the discovery of new antibacterial agents and illustrates the critical role played by genomics in each. The first approach is genomic target-based screening. Comparative genomics and bioinformatics are used to identify novel, selective ...

Genome sequencing, the determination of the complete complement of DNA in an organism, is revolutionizing all aspects of the biological sciences. Genome sequences make available for scientific scrutiny the complete genetic capacity of an organism. With respect to microbes, this mea ...

Microbial pathogens possess a repertoire of virulence determinants that make unique contributions to bacterial fitness during infection. In this chapter, we focus on the recent progress and adaptations of signature-tagged mutagenesis (STM) by PCR instead of hybridization. This ...

Recent advances in gene-amplification technology and molecular phylogenetics have provided the means of detecting and classifying bacteria directly from their natural habitats without the need for culture. These techniques have revolutionized environmental microbio ...

Isolates of bacterial species that are indistinguishable in genotype are assigned as a clone, with the implication that they are descended from the same recent ancestor. Clones are difficult to define with precision since bacteria are not truly asexual, and recombinational replaceme ...