微生物学技术

Mycobacterial cell wall ultrastructure has been studied through the use of negative staining, electron microscopy (1,2), freeze fracture (3), X-ray diffraction (4), differential scanning calorimetry (5,6), and electron spin resonance spectroscopy. Through the use of these techn ...

Great progress has been made in the latter half of the twentieth century in the understanding of the immunology of tuberculosis and of strategies for chemotherapeutic management of this disease. Indeed, given the evidence that the dominant, and perhaps sole, ecologic niche of Mycobacteri ...

Mycobacterium tuberculosis is a facultative intracellular pathogen, which can survive and replicate within the host macrophage. It is transmitted via the aerosol route, which delivers the bacillus to the alveolus of the lungs. The initial phase of granuloma formation within the lung re ...

The ability of pathogenic Mycobacterium to establish and maintain an infection in a host is dependent on their capacity to survive within phagocytes (1–3). Studies conducted on macrophage infections in culture have provided considerable insight into the mechanisms developed by the ...

Molecular beacons are a novel family of hybridization probes, which emit fluorescence upon interaction with their target. They are hairpin-shaped oligonucleotides with a central part complementary to the target, flanked by two 5 6 base pair (bp) inverted repeats, which can form a stable ste ...

Genetic manipulation of mycobacteria has historically been difficult. This is in large part due to the impenetrable nature of the cell wall, resulting in difficulty both in introducing DNA into the bacterium and subsequent isolation of intact plasmid DNA. In addition, the mycobacterial c ...

The complete genome sequence of the well studied laboratory strain of Mycobacterium tuberculosis (H37Rv) has been published by the Sanger Centre (1), and this information will soon be complemented by the completion of a recent clinical isolate of M. tuberculosis (CDC1551/CSU93) by The Ins ...

The term “proteomics” describes the technologies collectively used to define the protein complement of the genome or “proteome” (1,2). The recent growth of this discipline is reflected in the many review articles available (3–7). In addition to describing all the proteins encoded by the geno ...

Completion of the sequence of the entire genome of strain H37Rv was a benchmark for Mycobacterium tuberculosis research (1). This achievement ushers in the era of genome-wide functional and comparative genomics for this organism. At present, the most powerful enabling technology of the p ...

The storage and maintenance of mycobacterial reference strains and clinical isolates are important parts of good laboratory practice in a mycobacterial laboratory. The storage of reference and control strains facilitates a reliable control on intra- and inter-test reproducib ...

Research into and identification of Mycobacterium tuberculosis can take on a number of facets, many of which involve the use of DNA at one stage or another. The quality and quantity of DNA required will depend on the end-use requirement. For example, good yields of pure, high-molecular-weight DNA un ...

The worldwide resurgence of tuberculosis (TB) has resulted in a rapid expansion in research efforts directed at a deeper understanding of the disease, the efficacy of vaccines, and improved drug targets. This has required the establishment of multiple new facilities to contain the pathog ...

Pathogenicity in Mycobacterium tuberculosis may be thought of as a multifactorial process with both pathogen and host-response effector molecules contributing to the process of infection, leading either to immunopathology and disease or control of infection and long-term per ...

Genetic analyses of pathogenic mycobacteria such as Mycobacterium tubeculosis and Mycobacterium bovis required improvement of existing methodologies for the generation of large representative libraries of mutants. Two basic methodologies have been used to generate mut ...

Gene replacement and transposon mutagenesis are two complementary tools that have been widely used to perform genetic studies in various living organisms. In mycobacteria, and especially in the Mycobacterium tuberculosis complex, the lack of these tools has severely hampered the g ...

Much progress has been made in mycobacterial research in general and in mycobacterial genetics in particular during the past 10 yr. The complete genome sequences of two isolates of Mycobacterium tuberculosis, the widely distributed laboratory strain M. tuberculosis H37Rv (1) and a clin ...

Gene replacement by homologous recombination (HR) is an invaluable tool in understanding the physiology and the significance of specific genes in the virulence of Mycobacterium tuberculosis. It will also allow for the development of rationally attenuated strains as candidate vac ...

The production of cytokines can be controlled by the regulation of transcription, by the regulation of translation, and by post-translational mechanisms. Therefore, to understand better the control of cytokine production, it is important to measure both concentration of the free cyt ...

The multiprobe ribonuclease protection assay (RPA) is a highly sensitive and specific method for simultaneous detection and quantification of several species of mRNA. Three most distinct advantages of the multiprobe RPA method are: high sensitivity and specificity, capacity to si ...

Cytokines belong to a family of immunoregulatory peptide growth factors that are produced mainly by immune cells after immune challenge. Various cells of nonimmune cell origin such as epithelial and muscle cells are also capable of producing cytokines, as well as many tumor cells (1, 2). We have r ...