Method of detecting and counting microorganisms

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving viable micro-organism

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435 34, 435 32, 435 30, 435 29, 435 36, 435 38, 4352551, 43525422, 4352542, 435848, 435849, 435882, 435883, 435884, 435921, 435922, 435923, C12Q 106, C12Q 104, C12Q 118, C12Q 102

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057891915

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BRIEF SUMMARY
The present invention relates to a method for qualitatively detecting and quantitatively counting microorganisms, in particular those microorganisms which colonize the human skin. In addition, the present invention relates to agents for use in such methods.
Microorganisms are differentiated into prokaryotic microorganisms, that is those without any actual cell nucleus, and eukaryotic microorganisms, that is those having a cell nucleus.
The eukaryotes include algae, fungi and protozoa. The prokaryotes include bacteria. Bacteria are, in turn, classified into eubacteria, filament-forming bacteria, prosthecate and budding bacteria, actinomycetes, obligately parasitic bacteria, spirochaetes, cyanobacteria, archaebacteria and others.
Eubacteria are essentially subdivided into cocci, that is spherical bacteria (e.g. streptococci), essentially rod-shaped or elongate cylindrical non-sporulating bacteria (e.g. coryneform bacteria, propionibacteria, pseudomonads, enterobacteria, spore-forming rods (e.g. bacilli and clostridia) and curved rods (e.g. spirilla and vibrios).
In addition, the eubacteria can be subdivided, over and above the previously described groups, into Gram-positive and Gram-negative bacteria, that is bacteria whose cell wall is stained blue after the Gram test even after washing with alcohol, or those whose cell wall is washed colourless once again by the alcohol.
As a consequence of the structure of their cell walls, Gram-positive and Gram-negative bacteria possess differing properties; for example penicillin, which intervenes in the process of cell wall formation, principally has a lethal effect on Gram-positive bacteria (although on some Gram-negative bacteria as well).
Microorganisms in general, and bacteria in particular, are practically ubiquitous. For example, mycobacteria, streptococci, staphylococci and propionibacteria, in the main, can be found on healthy human skin. Coryneform bacteria, which are also present on the skin, have recently been identified as being responsible for the production of an unpleasant body odour due to the decomposition of apocrine sweat. included in the eukaryotes. Eukaryotes are living creatures whose cells (eucytes), in contrast to those of the so-called prokaryotes (procytes), possess a cell nucleus which is delimited from the remainder of the cytoplasm by means of a nuclear coat and nuclear membrane. The cell nucleus contains the hereditary information, which is stored in chromosomes.
Representatives of the mycobionts include, for example, yeasts (Protoascomycetes), moulds (Plectomycetes), mildew (Pyrenomycetes), downy mildew (Phycomycetes) and, of course, the upright fungi (Basidiomycetes).
While the fungi, including the Basidiomycetes, are not vegetable organisms, they do, like these, have a cell wall, vacuoles which are filled with cell sap, and a flow of plasm which is readily visible in the microscope. They do not contain any photosynthetic pigments and are C-heterotrophic. They grow under aerobic conditions and obtain energy by oxidizing organic substances. However, some representatives, for example yeasts, are facultative anaerobic organisms and are able to obtain energy by means of fermentation processes.
Dermatomycoses are diseases in which certain fungal species, in particular dermatophytes, penetrate the skin and hair follicles. Examples of the symptoms of dermatomycoses are blisters, exfoliation, rhagades and erosion, usually associated with itching or allergic eczema.
Dermatomycoses can essentially be subdivided into the following four groups: dermatophytoses (e.g. epidermophytosis, favus, microsporosis and trichophytosis), yeast mycoses (e.g. pityriasis, Candida infections, blastomycosis, Busse-Buschke disease, torulosis, piedra alba, torulopsidosis and trichosporosis), mould mycoses (e.g. aspergillosis, cephalosporidosis, phycomycosis and scopulariopsidosis), systemic mycoses (e.g. chromomycosis, coccidiomycosis and histoplasmosis).
The pathogenic or facultatively pathogenic organisms from the yeast group include Candida species (e.g. Candida al

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Larson et al, "J. Clinical Microbiology", pp. 604-608, Mar. 1986.

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