Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1996-06-25
1999-09-21
Jones, W. Gary
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 231, 536 243, 935 76, 935 77, 935 78, C12Q 168, C07H 2102, C07H 2104, C12N 1500
Patent
active
059552670
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to the detection and characterization of yeasts belonging to the species Candida krusei. In particular, it provides a specific probe, especially a nucleic acid probe, for the detection and particularly identification of the species Candida krusei and for the typing (infrapecific characterization) of strains of the species Candida krusei.
PRIOR ART
Yeasts are fungi, i.e. eukaryotic microorganisms, in which the unicellular form is predominant (Barnett J. W., R. W. Payne and D. Yarrow, Yeasts: characteristics and identification, 2nd Edition, Cambridge University Press, Cambridge, 1991).
The genus Candida is a heterogeneous genus which groups together anamorphous species, i.e. species whose mode of sexual reproduction is or was unknown. When this mode of sexual reproduction is discovered, the species are generally reclassified within an already described species (teleomorph). Thus Issatchenkia orientalis is the teleomorph of Candida krusei.
In the taxonomy of yeasts, it is therefore possible to encounter two different species names for describing one and the same taxon (group) of yeasts.
In normal practice, the species name Candida krusei is the most commonly used and it is the one which will be employed indiscriminately in the present disclosure to denote Candida krusei or Issatchenkia orientalis.
Candida krusei is one of the ten principal species of yeasts of the genus Candida which are responsible for infections in humans or animals. This species is sometimes responsible for the degradation of food products. It is also used for certain industrial biotransformations.
Candida krusei infections are called "C. krusei candidoses" or "C. krusei mycoses"; they can affect practically any tissues in the human body. Systemic, generalized or deep infections are the most serious and can be fatal. The mortality rate can be high.
Yeasts of the species Candida krusei are opportunist and ubiquitous; their prevalence and pathogenicity are high in immunosuppressed subjects, especially in patients suffering from neutropenia, AIDS, cancer etc., who are particularly sensitive to C. krusei infections.
Finally, C. krusei candidoses arc obviously very contagious in the same way as any other fungal diseases.
The whole difficulty of treating infections of this type arises from the fact that strains of Candida krusei are relatively resistant to certain antifungal agents and to fluconazole in particular. What is even more troublesome, however, is that the use of these antifungal agents in prophylaxis, or even in therapy, would favor colonization of the subjects by endogenous strains if they were present in the patient.
Consequently, rapid detection and precise and reliable identification, as well as typing of the strains of this species, are increasingly necessary, especially for enabling an appropriate antifungal treatment to be applied. Moreover, there is a real need for reliable epidermiological markers for use in epidermiological studies or for tracing stains in the agri-foodstuffs or fermentation industries.
One is forced to note, however, that the conventional methods of identifying yeasts are not entirely satisfactory in all these respects.
Thus, known methods include especially those described by Barnett, J. W., R. W. Payne and D. Yarrow in: "Yeasts: characteristics and identification", 2nd edition, Cambridge University Press, Cambridge, 1991, which require prior isolation of the microorganism in pure culture and then a study of its morphological and especially physiological characteristics (more than 80 tests), these ions together taking from 10 to 30 days for one identification. These methods are therefore tedious and particularly long. They are restricted to reference laboratories and cannot be used in normal practice.
Although more rapid and more rational, the miniaturized and standardized methods of identifying yeasts of medical importance still include prior isolation and then identification of the microorganism by meals of physiological tests, which are performed for exa
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Carlotti Arnaud
Villard Jean
Bio Merieux S.A.
Jones W. Gary
Whisenant Ethan
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