微生物学技术

Mycoplasmas are parasites and pathogens of plants, insects, and animals, including humans. Although mycoplasmas are highly evolved parasites, they are not immune to being parasitized themselves. Within the class Mollicutes, extrachromosomal elements have been described for s ...

Transposon mutagenesis, although utilized extensively in cell-walled bacteria, has been used rarely within the class Mollicutes (1). The reasons for such limited use include a lack of the necessary techniques and transposons that contain suitable antibiotic markers. There has been l ...

Transformation of mycoplasmas was not clearly demonstrated until 1987 when Dybvig and Cassell were able to show the introduction of the Gram-positive transposon Tn916 into Acholeplasma laidlawii and Mycoplasma pulmonis (1). This was the first direct evidence that mycoplasmas cou ...

Workers using cell cultures for research, diagnostics, or production of biopharmaceuticals have in common the need to maintain their cultures in a state of control. The living cell-culture system that is expected to produce reliably scientific and diagnostic data or a product must not be enc ...

Prokaryotic insertion sequences (IS) are transposable genetic elements with a length ranging from 800–2500 bp. They are found at a remarkable variety in the genomes of many different bacteria and mycoplasmas at a multiplicity of between a few and several hundred per genome. In bacteria, they a ...

In routine diagnosis, bacterial field isolates are identified by serological and/or biochemical tests. Occasionally, however, isolates deviate from the generally accepted patterns, i.e., they may react with diagnostic antisera of more than one type strain or fail to react with any of them, ...

The diagnosis and typing of microorganisms of human and veterinary significance are vitally important in the rapid and effective treatment of infectious diseases. Traditionally, the growth of bacteria in specific media and/or the immunological detection of surface antigens is us ...

The reappearance of contagious bovine pleuropneumonia (CBPP) in Italy in 1990 after an absence of 100 years and the continued spread of CBPP in many parts of Africa are powerful reminders that this age-old disease is still a long way from eradication. Control of this disease was almost achieved dur ...

The polymerase chain reaction (PCR), DNA hybridization, and sequence analysis have been valuable in the study of the phylogenic relationships between members of the Mycoplasma mycoides “cluster” (1). They have confirmed the very close relationships between these organisms sugges ...

Ribosomes are present in all self-replicating cells and constitute their protein-synthesizing machinery. The ribosomes are composed of ribosomal proteins and ribosomal RNA (rRNA). Bacteria have three kinds of rRNA (5S, 16S, and 23S rRNA), and the genetic information of these molecules ...

The manipulation of genetic material for the purpose of diagnosis or analysis almost always requires the preparation of sample to expose genomic nucleic acid or the extraction and purification of DNA. Molecular techniques, such as restriction enzyme analysis, Southern hybridizat ...

Immunohistochemistry is a technique in which the specific interaction between an immunoglobulin and its homologous antigen is visualized on histological sections by a microscopically detectable label. Generally, the label consists of an enzyme, such as peroxidase, alkaline ph ...

The advantages of microtiter-based ELISAs in diagnostic techniques can be briefly summarized by the economic use of reagents and by the ease of their application to large numbers of test samples. ELISAs are widely applied to the serological diagnosis of both human and animal bacterial and myc ...

Immunofluorescence has been used as a diagnostic tool for identification of mycoplasmas cultured on artificial medium and also for detection of the organisms in situ in infected hosts and for detection of contaminated cell cultures. The technique has been used in research investigati ...

The definitive identification of mycoplasmas is usually based on serological procedures, including growth inhibition (1,2), metabolic inhibition (3,4), immunofluorescence (5–7), and immunobinding assays (8–14). The technique most commonly adopted for the routine identif ...

The most common application of the polymerase chain reaction (PCR) IS to exponenttally amplify a specific known and predictable sequence from a complex mixture of nucleic acids. This chapter describes a technique that, in contrast, uses PCR with arbitrary primers, to generate a fingerpri ...

Tetracyclines are antimicrobial agents that interact with bacterial ribosomes and block protein synthesis. They have activity against a wide range of gram positive, gram-negative, anaerobic and aerobic bacteria, cell-wall free myco plasmas, chlamydiae, mycobacteria, ricke ...

The polymerase chain reaction (PCR) was first described in 1985 (1), although its theoretical roots go back beyond that time (2). It is the most versatile of the amplification methods, the others (see Subheading 4.) are more or less confined to diagnostic applications. For example, the product of a PCR, ...

Quinolone antibacterial agents were first introduced into the clinical environment in the early 1960s. The first qumolone to be clinically used was nalidixic acid, which was used for the treatment of enteric and urinary tract infections. As a result of increased clinical resistance to this ...

Our original intention was for this chapter to be written by Sandra Handwerger (Rockefeller University, New York), but tragically she died in April 1996 before she could undertake the task. In our opinion, Sandra was one of the leading research workers on genetic aspects of glycopeptide resist ...