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2008 | 64 | 06 | 822-827
Tytuł artykułu

Wewnatrzgatunkowe roznicowanie szczepow Malassezia pachydermatis z wykorzystaniem PCR-REA

Warianty tytułu
EN
Intraspecies differentiation of Malassezia pachydermatis strains with PCR-REA technique
Języki publikacji
PL
Abstrakty
EN
The aim of the study was to make an attempt at showing the intraspecies heterogenecity of Malassezia pachydermatis strains with regards to their origin (strains isolated from healthy dogs and with otitis externa symptoms). The study included 41 strains of Malassezia pachydermatis species isolated in a pure culture from dogs with clinical otitis externa symptoms (n = 20), clinically healthy dogs (n = 20) and a reference strain, M. pachydermatis (CBS7925). In order to isolate the genetic material from the fungal cells, the following four procedures were selected: mechanical, enzymatic, thermal and chemical. Considering the yield and repeatability of a method for the genomic DNA extraction, a mechanical method was applied. The genetic material research of each strain was performed according to PCR-REA technique with the amplification of three genome regions: ITS, LSU rRNA and a gene encoding beta-tubuline. The ITS and LSU rRNA regions were amplified employing the standard PCR reagents, whereas the region coding beta-tubuline with the so called touch down. The obtained amplification products were subjected to restrictive analysis by means of the following enzymes: EcoRI, Ncol, Hinfl, Alul, and Eco881 (Aval). The performed investigations made it possible to reveal the genotypic differentiation within M.pachydermatis species as well as some correlation between a genotypic profile and the origin of a strain (from healthy animals or with otitis externa symptoms), which may imply the existence of genetic conditioning of the Malassezia strains’ pathogenicity.
Wydawca
-
Rocznik
Tom
64
Numer
06
Strony
822-827
Opis fizyczny
s.822-827,fot.,tab.,bibliogr.
Twórcy
  • Uniwersytet Przyrodniczy w Lublinie, ul.Akademicka 12, 20-033 Lublin
Bibliografia
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  • 3. David M., Gabriel M., Kopecka M.: Unusual ultrastructural characteristic of yeast Malassezia pachydermatis. Scripta Medicina 2003, 76, 173-186.
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  • 6. Gil-Lamaigniere C., Roilidies E., Hacher J., Muller F. M. C.: Molecular typing for fungi - a critical review of the possibilities and limitations of currently and future methods. Eur. Soc. Clin. Microbiol. Inf. Dis. 2003, 9, 172-185.
  • 7. Guillot J.: Epidemiological analysis of Malassezia pachydermatis isolates by partial sequencing of the large subunit ribosomal RNA. Res. Vet. Sci. 1997, 62, 22-25.
  • 8. Guillot J., Deville M., Berthelemy M., Provost F., Gueho E.: A single PCR-restriction endonuclease analysis for rapid identification of Malassezia species. Lett. Appl. Microbiol. 2000, 31, 400-403.
  • 9. Guillot J., Gueho E.: The diversity of Malassezia yeasts confirmed by rRNA sequence and nuclear DNA comparisons. Antonie van Leeuwenhoek. 1995, 67, 297-314.
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  • 11. Gupta A. K., Kohli Y., Summerbell R. C.: Molecular differentiation of seven Malassezia species. J. Clin. Microbiol. 2000, 38, 1869-1875.
  • 12. Gursharan S., Walker M. J., Yan H., Timnis K. N., Guzman C. A.: Temperature dependent expession of an acid phosphatase by Bordetella bronchiseptica: role in intracellular survival. Microb. Pathogenesis 1997, 22, 257-164.
  • 13. Inamader A. C., Palit A.: The genus Malassezia and human disease. Indian J. Dermatol. 2003, 69, 265-270.
  • 14. Makimura K., Tamura Y., Kudo M., Uchida K., Saito H., Yamaguchi H.: Species identification and strain typing of Malassezia species stock strains and clinical isolates based on the DNA sequences of nuclear ribosomal internal transcribed spacer 1 regions. J. Med. Microbiol. 2000, 49, 29-35.
  • 15. Nowakiewicz A.: Współczesne metody różnicowania diagnostycznego szczepów z rodzaju Malassezia izolowanych z przypadków klinicznych. Rozprawa dokt. Wydz. Medycyny Weterynaryjnej, AR Lublin 2005.
  • 16. Nowakiewicz A., Ziółkowska G.: Fenotypowy profil patogenności szczepów Malassezia pachydermatis. Medycyna Wet. 2007, 63, 721-727.
  • 17. Sambrook J., Fritsch E. F., Maniatis T.: Molecular Cloning (A laboratory manual). Cold Spring Harbor Laboratory Press, New York 1989.
  • 18. Senczek D., Siesenop U., Bohm K. H.: Characterization of Malassezia species by means of phenotypic characteristics and detection of electrophoretic karyotypes by pulsed-field gel electrophoresis (PFGE). Mycoses 1999, 42, 409-414.
  • 19. Shattuck K. E., Cochran C. K., Zabrausky R. J., Pasarell L., Davis J. C., Malloy M. H.: Colonization and infection associated with Malassezia and Candida species in a neonatal unit. J. Hosp. Infect. 1996, 34, 123-129.
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  • 22. Sugita T., Takeo K., Hama K., Virtudazo E., Takashima M., Nishikawa A., Kucsera J., Dorogi J., Komori S., Nakagaki K., Vollekova A., Slavikova E., Farka F.: DNA sequence diversity of intergenic spacer I region in the non-lipid-dependent species Malassezia pachydermatis isolated from animals. Med. Mycol. 2005, 43, 21-26.
  • 23. Van Burik J., Myerson D., Schreckhise R. W., Bowden R. A.: Panfungal PCR assay for detection of fungal infection in human blood specimens. J. Clin. Microbiol. 1998, 36, 1169-1175.
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Typ dokumentu
Bibliografia
Identyfikatory
Identyfikator YADDA
bwmeta1.element.agro-article-66fe1505-24fc-4328-997f-ab2e3f991117
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