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The Detection of Enteroviruses in Water and Associated Materials Using the Polymerase Chain Reaction

The environment furnishes a variety of surroundings that may favor the survival of microorganisms. The aquatic environment (which includes marine and fresh water, raw and treated sewage, sludge, and sediments) often provides conditions in which pathogenic viruses released from the ...

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Robotic Analyzers

In general, three assay types are commonly used in diagnostic virology. Since these are described in other chapters it suffices here just to reiterate the methodology names: 1.

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Recombinant Antigens in Viral Diagnosis

Traditionally, the accurate detection of viruses and the diagnosis of viral diseases has been difficult and expensive as viruses cannot be visualized by conventional light microscopy and need propagation in primary or continuous cell culture. The advent and increasing sophistica ...

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Oligonucleotides

During the past 30 years, several strategies have been used to synthesize DNA. Moreover, it was not possible to utilize automated DNA synthesis until the phosphate triester method using phosphoramidite reagents was introduced (1,2). Since then, chemically synthesized oligonucleo ...

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NASBA: A Method for Nucleic Acid Diagnostics

Nucleic acid sequence-based amplification (NASBA) is a primer dependent, homogeneous, isothermal amplification process for the detection of RNA (1). It can result in 109- fold amplification of a specific RNA sequence and offers the unique possibility to amplify RNA in a background of geno ...

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Enzyme Analysis: The Rationale and Use of Enzyme Assays in Assigning Function to Gene Nucleotide Sequences and the Procedures for the Assay of Three E

An issue of some biological currency pertains to the identification of the functional potential of cells as deduced from the nucleotide sequence of their genes. In effect, nucleotide sequences of genes assigned in some confidence to expressed proteins and function in one cell are taken, when ...

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Detection of Mycoplasmas in Cell Culture by Fluorescence Methods

The fluorescence stains used most often for mycoplasma detection are DNA binding fluorochromes (DNAFs) and fluoresceinated antibodies. Both permit direct visualization of individual mycoplasma organisms. DNAFs will bind to any appropriately conformed DNA that is present in a s ...

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Serological Identification of Mycoplasmas by Growth and Metabolism Inhibition Tests

The lack of a cell wall means that the growth of mycoplasmas can be inhibited by specific antiserum and provides the basis of a simple, economic, and objective means of species identification. In the diagnostic laboratory, unknown mycoplasma isolates are inoculated onto solid media and exami ...

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Molecular Approaches for the Detection and Identification of -Lactamases

β-lactamases confer resistance to β-lactam antibiotics, which are the most widely used family of antibiotics. It is, therefore, essential that one can identify the production of β-lactamases by clinical isolates and have effective ways of distinguishing the different enzymes. This is n ...

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Application of Denaturing Gradient Gel Electrophoresis to Microbial Ecology

A significant proportion of microbial ecology is now based on the description of community structure in naturally occurring bacterial assemblages. The development of molecular biological techniques has facilitated this task, primarily via the cloning and sequencing of microb ...

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Recovery and Analysis of Ribosomal RNA Sequences from the Environment

Since the 1980s the use of ribosomal RNA (rRNA) sequence-based analysis to characterize microbial populations (mainly bacterial and archaeal populations) has increased significantly. This increased use is in response to the recognition that culture-based methods grossly mis ...

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Fluorescent Polymerase Chain Reaction/Restriction Fragment Length Polymorphism Monitoring of Genes Amplified Directly from Bacterial Communities in So

There has been a growing acknowledgment of both the ecological and biotechnological importance of microbes in natural environments. Concerns about the nonrepresentative nature of traditional analytical methods, as a result of their requirement for prior cultivation, have led to ...

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Confocal Laser Scanning Microscopy of Environmental Samples

Enormous technical advances in imaging and data acquisition techniques, combined with a continuing increased scope for fluorescence labeling of specific constituents of living organisms, have brought about a revolution in approaches to biological problems. Whereas spatial ...

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Fluorescent In Situ Hybridization and the Analysis of the Single Cell

The discovery that prokaryotic and eukaryotic cells could be made permeable to fluorescently labeled, sequence specific oligonucleotides makes possible the determinative probing of intact microbial cells (1). Thus, individual target cells can be identified and enumerated in h ...

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Characterizing Microorganisms in the Environment by Fatty Acid Analysis

Determining the taxonomic composition, biomass, and physiological status of microbial assemblages is still one of the greatest challenges facing microbial ecologists. There are many reasons why assessment of microbes in the environment is so demanding, not least their number, div ...

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Specific Detection, Viability Assessment, and Macromolecular Staining of Bacteria for Flow Cytometry

Direct analysis of bacteria from natural environments is problematic. Direct examination of samples by microscopy is an essential technique for bacteriologists, but is prone to error, is time-consuming, and can be tedious. In many situations, the process can be automated using flow cyto ...

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Analysis of DNA Sequences

Molecular biological methods are now commonly used to detect bacteria in diverse environments ranging from soils, sediments, and sludges (1,2) to plant (3) and mammalian (4,5) tissue and food or water samples (6,7). The techniques most widely used in detection methods are the polymerase chain r ...

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Automated Sequencing of DNA Retrieved from Environmental Samples

The advent of molecular techniques has revolutionized our understanding of microbial ecology, and their use in environmental microbiology is widespread. Many of the recent studies investigating the microbial flora of diverse ecosystems have adopted a common approach of targeted ...

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DNA Extraction from Natural Environments

Until recently, studies on microbial communities present in natural environments relied on conventional optical microscopic observation and cultivation-based approaches. Although these traditional approaches remain valuable, they have a number of limitations. The mo ...

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Magnetic Particle-Based Separation Techniques for Monitoring Bacteria from Natural Environments

Physical separation of either intact target cells or specific molecules from many environments can result in a suspension free of contaminating particles, nontarget cells and biological inhibitors and highly enriched in the target cells or molecules of interest. The processed samp ...

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