实验基础

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 ...

Aminoglycosides have been cinically used since 1944. Although this class of antibacterial agents has some nephrotoxicity and ototoxtcity issues, they continue to be part of the hospital armamentarium because of then rapid bactericidal activity, especially in combination with β- ...

The widespread use, or perhaps overuse, of penicillin during the past 50 yr has driven the evolution of resistance to penicilling in numerous different species of bacteria.Typically, resistance has arisen as a result of the acquisition of β-lactamases that inactivate the antibiotic (see ...

For the last 20 yr thin-layer polyacrylamide isoelectric focusing (IEF) has played a major role in the identification and characterization of β-lactamases. IEF is able to distinguish enzymes that focus only 0.05 pI apart (1), but the exponential increase rin the numbers of β-lactamases disco ...

Bacterial plasmids are extra-chromosomal, covalently-closed circular (CCC) molecules of DNA that are capable of autonomous replication (1). Plasmids may contain genes for a variety of phenotypic traits, such as antibiotic resistance, virulence, or metabolic activities, altho ...

Enterococci are components of the normal bowel flora of humans and other animals, and have traditionally been considered to be of relatively low virulence in healthy individuals. However, they are increasingly important nosocomial pathogens and have been cited as the leading organism ...

Staphylococcus aureus and coagulase-negative species of staphylococci (CNS), particularly S epidermidis, are among the most frequently isolated bacteria from patients with nosocomial infection. Conventional methods used in the clinical microbiology laboratory for id ...

Helicobacter pylori is a curved mlcroaerobic bacterium that was first lsolated from human antral gastric biopsy material in 1982 by Marshall and colleagues in Perth, Western Australia (1). Since then, enormous interest has developed in the micro-organism that now appears to be one of the most ...