1887

Abstract

and are recently described species phenotypically indistinguishable from . We evaluated phenotyping and molecular methods for the detection of these species among 79 unique blood culture isolates of the group obtained during the years 2004–2008. The isolates were screened by PCR amplification of the secondary alcohol dehydrogenase-encoding gene () followed by digestion with the restriction enzyme I, using ATCC 22019, ATCC 96139 and ATCC 96144 as controls. Isolates with RFLP patterns distinct from were characterized by sequence analysis of the ITS1–ITS2, 26S rRNA (D1/D2) and regions. Restriction patterns for the 3 species with each of 610 restriction enzymes were predicted using 12 available sequences. By PCR-RFLP of the gene alone, four isolates (5.1 %) had a pattern identical to the reference strain. Sequence analysis of and ITS (internal transcribed spacer) regions identified two of these isolates as . These results were confirmed by creating a phylogenetic tree based on concatenated sequences of , ITS and 26S rRNA gene sequence regions. Optimal differentiation between , and was predicted using digestion with III, producing discriminatory band sizes of: 131 and 505 bp; 74, 288 and 348 bp; and 131, 217 and 288 bp, respectively. This was confirmed using the reference strains and 79 clinical isolates. In conclusion, reliable discrimination was obtained by PCR-RFLP profile analysis of the gene after digestion with III but not with I. and are involved in a small but significant number of invasive infections in Denmark.

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2010-04-01
2019-10-13
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References

  1. AFST-EUCAST ( 2008; ). EUCAST definitivedocument EDef 7.1: method for the determination of broth dilution MICs ofantifungal agents for fermentative yeasts. Clin Microbiol Infect 14, 398–405.[CrossRef]
    [Google Scholar]
  2. Arendrup, M. C., Fuursted, K., Gahrn-Hansen, B., Schønheyder,H. C., Knudsen, J. D., Jensen, I. M., Bruun, B., Christensen, J. J. &Johansen, H. K. ( 2008; ). Semi-national surveillanceof fungaemia in Denmark 2004–2006: increasing incidence of fungaemiaand numbers of isolates with reduced azole susceptibility. ClinMicrobiol Infect 14, 487–494.
    [Google Scholar]
  3. Asadzadeh, M., Ahmad, S., Al-Sweih, N. & Khan, Z. U. ( 2009; ). Rapid molecular differentiation and genotypic heterogeneityamong Candida parapsilosis and Candida orthopsilosis strainsisolated from clinical specimens in Kuwait. J Med Microbiol 58, 745–752.[CrossRef]
    [Google Scholar]
  4. Borman, A. M., Linton, C. J., Oliver, D., Palmer, M. D., Szekely,A., Odds, F. C. & Johnson, E. M. ( 2009; ). Pyrosequencinganalysis of 20 nucleotides of internal transcribed spacer 2 discriminates Candida parapsilosis, Candida metapsilosis, and Candidaorthopsilosis. J Clin Microbiol 47, 2307–2310.[CrossRef]
    [Google Scholar]
  5. Campa, D., Tavanti, A., Gemignani, F., Mogavero, C. S., Bellini,I., Bottari, F., Barale, R., Landi, S. & Senesi, S. ( 2008; ). DNA microarray based on arrayed-primer extension technique foridentification of pathogenic fungi responsible for invasive and superficialmycoses. J Clin Microbiol 46, 909–915.[CrossRef]
    [Google Scholar]
  6. Enger, L., Joly, S., Pujol, C., Simonson, P., Pfaller, M. A. &Soll, D. R. ( 2001; ). Cloning and characterization ofa complex DNA fingerprinting probe for Candida parapsilosis. J Clin Microbiol 39, 658–669.[CrossRef]
    [Google Scholar]
  7. Fell, J. W. ( 1993; ). Rapid identificationof yeast species using three primers in a polymerase chain reaction. Mol Mar Biol Biotechnol 2, 174–180.
    [Google Scholar]
  8. Garcia-Effron, G., Katiyar, S. K., Park, S., Edlind, T. D. &Perlin, D. S. ( 2008; ). A naturally occurring proline-to-alanineamino acid change in Fks1p in Candida parapsilosis, Candida orthopsilosis, and Candida metapsilosis accounts for reduced echinocandinsusceptibility. Antimicrob Agents Chemother 52, 2305–2312.[CrossRef]
    [Google Scholar]
  9. Gomez-Lopez, A., Alastruey-Izquierdo, A., Rodriguez, D., Almirante,B., Pahissa, A., Rodriguez-Tudela, J. L., Cuenca-Estrella, M. & the BarcelonaCandidemia Project Study Group ( 2008; ). Prevalenceand susceptibility profile of Candida metapsilosis and Candidaorthopsilosis: results from population-based surveillance of candidemiain Spain. Antimicrob Agents Chemother 52, 1506–1509.[CrossRef]
    [Google Scholar]
  10. Gonçalves, S. S., Amorim, C. S., Nucci, M., Padovan,A. C. B., Briones, M. R. S., Melo, A. S. A. & Colombo, A. L. ( 2010; ). Prevalence rates and antifungal susceptibility profilesof C. parapsilosis species complex: results from a nationwide surveillanceof candidaemia in Brazil. Clin Microbiol Infect 16 (in press)
    [Google Scholar]
  11. Hensgens, L. A., Tavanti, A., Mogavero, S., Ghelardi, E. &Senesi, S. ( 2009; ). AFLP genotyping of Candidametapsilosis clinical isolates: evidence for recombination. Fungal Genet Biol 46, 750–758.[CrossRef]
    [Google Scholar]
  12. Iwen, P. C., Hinrichs, S. H. & Rupp, M. E. ( 2002; ). Utilization of the internal transcribed spacer regions asmolecular targets to detect and identify human fungal pathogens. Med Mycol 40, 87–109.[CrossRef]
    [Google Scholar]
  13. Kato, M., Ozeki, M., Kikuchi, A. & Kanbe, T. ( 2001; ). Phylogenetic relationship and mode of evolution of yeastDNA topisomerase II gene in the pathogenic Candida species. Gene 272, 275–281.[CrossRef]
    [Google Scholar]
  14. Kocsubé, S., Tóth, M., Vágvölgyi,C., Dóczi, I., Pesti, M., Pócsi, I., Szabó, J. &Varga, J. ( 2007; ). Occurrence and genetic variabilityof Candida parapsilosis sensu lato in Hungary. J Med Microbiol 56, 190–195.[CrossRef]
    [Google Scholar]
  15. Krcmery, V. & Barnes, A. J. ( 2002; ). Non-albicans Candida spp. causing fungaemia: pathogenicity andantifungal resistance. J Hosp Infect 50, 243–260.[CrossRef]
    [Google Scholar]
  16. Lehmann, P. F., Lin, D. M. & Lasker, B. A. ( 1992; ). Genotypic identification and characterization of speciesand strains within the genus Candida by using random amplified polymorphicDNA. J Clin Microbiol 30, 3249–3254.
    [Google Scholar]
  17. Lin, D., Wu, L. C., Rinaldi, M. G. & Lehmann, P. F. ( 1995; ). Three distinct genotypes within Candida parapsilosis from clinical sources. J Clin Microbiol 33, 1815–1821.
    [Google Scholar]
  18. Lockhart, S. R., Messer, S. A., Pfaller, M. A. & Diekema,D. J. ( 2008; ). Geographic distribution and antifungalsusceptibility of the newly described species Candida orthopsilosisand Candida metapsilosis in comparison to the closely related species Candida parapsilosis. J Clin Microbiol 46, 2659–2664.[CrossRef]
    [Google Scholar]
  19. Lott, T. J., Kuykendall, R. J., Welbel, S. F., Pramanik, A. &Lasker, B. A. ( 1993; ). Genomic heterogeneity in theyeast Candida parapsilosis. Curr Genet 23, 463–467.[CrossRef]
    [Google Scholar]
  20. Mirhendi, H., Diba, K., Rezaei, A., Jalalizand, N., Hosseinpur,L. & Khodadadi, H. ( 2007; ). Colony-PCR is a rapidand sensitive method for DNA amplification in yeasts. Iran J PublicHealth 36, 40–44.
    [Google Scholar]
  21. Nosek, J., Tomaska, L., Rycovska, A. & Fukuhara, H. ( 2002; ). Mitochondrial telomeres as molecular markers foridentification of the opportunistic yeast pathogen Candida parapsilosis. J Clin Microbiol 40, 1283–1289.[CrossRef]
    [Google Scholar]
  22. Pryce, T. M., Palladino, S., Price, D. M., Gardam, D. J., Campbell,P. B., Christiansen, K. J. & Murray, R. J. ( 2006; ). Rapid identification of fungal pathogens in BacT/ALERT, BACTEC, andBBL MGIT media using polymerase chain reaction and DNA sequencing of the internaltranscribed spacer regions. Diagn Microbiol Infect Dis 54, 289–297.[CrossRef]
    [Google Scholar]
  23. Roy, B. & Meyer, S. A. ( 1998; ). Confirmationof the distinct genotype groups within the form species Candida parapsilosis. J Clin Microbiol 36, 216–218.
    [Google Scholar]
  24. Silva, A. P., Miranda, I. M., Lisboa, C., Pina-Vaz, C. &Rodrigues, A. G. ( 2009; ). Prevalence, distributionand antifungal susceptibility profile of Candida parapsilosis, C. orthopsilosis and C. metapsilosis in a tertiary care hospital. J Clin Microbiol 47, 2392–2397.[CrossRef]
    [Google Scholar]
  25. Tavanti, A., Davidson, A. D., Gow, N. A., Maiden, M. C. &Odds, F. C. ( 2005; ). Candida orthopsilosisand Candida metapsilosis spp. nov. to replace Candida parapsilosis groups II and III. J Clin Microbiol 43, 284–292.[CrossRef]
    [Google Scholar]
  26. Tavanti, A., Hensgens, L. A. M., Ghelardi, E., Campo, M. &Senesi, S. ( 2007; ). Genotyping of Candida orthopsilosis clinical isolates by amplification fragment length polymorphism revealsgenetic diversity among independent isolates and strain maintenance withinpatients. J Clin Microbiol 45, 1455–1462.[CrossRef]
    [Google Scholar]
  27. Tay, S. T., Na, S. L. & Chong, J. ( 2009; ). Molecular differentiation and antifungal susceptibilities of Candidaparapsilosis isolated from patients with bloodstream infections. J Med Microbiol 58, 185–191.[CrossRef]
    [Google Scholar]
  28. van Asbeck, E., Clemons, K. V., Martinez, M., Tong, A. J. &Stevens, D. A. ( 2008; ). Significant differences indrug susceptibility among species in the Candida parapsilosis group. Diagn Microbiol Infect Dis 62, 106–109.[CrossRef]
    [Google Scholar]
  29. White, T. J., Bruns, T. D., Lee, S. B. & Taylor, J. W. ( 1990; ). Amplification and direct sequencing of fungal ribosomalRNA genes for phylogenetics. In PCR Protocols: a Guide to Methods andApplications, pp. 315–322. Edited by M. A. Innis, D. H. Gelfand,J. J. Sninsky & T. J. White. San Diego, CA: Academic Press.
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