1887

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Volume 58, Issue 6

Rapid molecular differentiation and genotypic heterogeneity among and strains isolated from clinical specimens in Kuwait Free

Abstract

Recent molecular studies have led to the recognition of three distinct species within the complex, namely , and . As currently available yeast identification systems fail to differentiate these species, there is a paucity of information on their occurrence in different geographical regions. This study describes a simple PCR-based protocol for rapid discrimination among , and strains by using primers derived from unique sequences within the internally transcribed spacer 1 (ITS1)–5.8 rRNA–ITS2 region. Retrospective analysis of 114 -complex isolates recovered from clinical specimens in Kuwait identified 109 as , five as and none as . The results were further validated by PCR-RFLP patterns of the secondary alcohol dehydrogenase gene fragment. DNA sequencing of the ITS region and the D1/D2 regions of the 28S rRNA gene confirmed the species-specific identification of all five strains. The amplicon length of the intergenic spacer between the 28S and 5S rRNA genes (IGS1) was also species-specific, and PCR-RFLP analyses of the IGS1 region identified two distinct genotypes among the five strains, which corresponded with the ITS region sequence data. The three bloodstream strains were confined to a single genotype. Among 81 randomly selected strains, two genotypes were detected by IGS1 region analyses, indicating limited genotypic heterogeneity among strains. As far as is known, this is the first report on the identification of from a bloodstream infection in the Arabian Gulf region.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): IGS, intergenic spacer and ITS, internally transcribed spacer
SGM
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2009-06-01
2024-04-24
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References

  1. Ahmad S., Khan Z., Mustafa A. S., Khan Z. U. 2002; Seminested PCR for diagnosis of candidemia: comparison with culture, antigen detection, and biochemical methods for species identification. J Clin Microbiol 40:2483–2489 [CrossRef]
     [Google Scholar]
  2. Ahmad S., Mustafa A. S., Khan Z., Al-Rifaiy A. I., Khan Z. U. 2004; PCR-enzyme immunoassay of rDNA in the diagnosis of candidemia and comparison with amplicon detection by agarose gel electrophoresis. Int J Med Microbiol 294:45–51 [CrossRef]
     [Google Scholar]
  3. Ahmad S., Al-Mahmeed M., Khan Z. U. 2005; Characterization of Trichosporon species isolated from clinical specimens in Kuwait. J Med Microbiol 54:639–646 [CrossRef]
     [Google Scholar]
  4. Ahmad S., Jaber A.-A., Mokaddas E. 2007; Frequency of embB codon 306 mutations in ethambutol-susceptible and -resistant clinical Mycobacterium tuberculosis isolates in Kuwait. Tuberculosis (Edinb 87:123–129 [CrossRef]
     [Google Scholar]
  5. Almirante B., Rodriguez D., Cuenca-Estrella M., Almela M., Sanchez F., Ayata J., Alonso-Tarres C., Rodriguez-Tudela J. L., Pahissa A. 2006; Epidemiology, risk factors, and prognosis of Candida parapsilosis bloodstream infections: case–control population-based surveillance study of patients in Barcelona, Spain, from 2002 to 2003. J Clin Microbiol 44:1681–1685 [CrossRef]
     [Google Scholar]
  6. Al-Obaid I., Ahmad S., Khan Z. U., Dinesh B., Hejab H. M. 2006; Catheter-associated fungemia due to Exophiala oligosperma in a leukemic child and review of fungemia cases caused by Exophiala species. Eur J Clin Microbiol Infect Dis 25:729–732 [CrossRef]
     [Google Scholar]
  7. Al-Sweih N., Ahmad S., Khan Z. U., Khan S., Chandy R. 2005; Prevalence of Candida dubliniensis among germ tube-positive Candida isolates in a maternity hospital in Kuwait. Mycoses 48:347–351 [CrossRef]
     [Google Scholar]
  8. Al-Sweih N., Khan Z. U., Khan S., Devarajan L. 2009; Neonatal candidaemia in Kuwait: a 12-year study of risk factors, species spectrum and antifungal susceptibility. Mycoses 52: (in press). doi: 10.1111/j.1439-0507.2008.01637
    [Google Scholar]
  9. Asadzadeh M., Al-Sweih N. A., Ahmad S., Khan Z. U. 2008; Antifungal susceptibility of clinical Candida parapsilosis isolates in Kuwait. Mycoses 51:318–323 [CrossRef]
     [Google Scholar]
  10. Bassetti M., Righi E., Costa A., Fasce R., Molinari M. P., Rosso R., Pallavicini F. B., Viscoli C. 2006; Epidemiological trends in candidemia in intensive care. BMC Infect Dis 6:21 [CrossRef]
     [Google Scholar]
  11. Bonassoli L. A., Bertoli M., Svidzinski T. I. 2005; High frequency of Candida parapsilosis on the hands of healthy hosts. J Hosp Infect 59:159–162 [CrossRef]
     [Google Scholar]
  12. Gee S. F., Joly S., Soll D. R., Meis J. F., Verweij P. E., Polacheck I., Sullivan J. D., Coleman D. C. 2002; Identification of four distinct genotypes of Candida dubliniensis and detection of microevolution in vitro and in vivo. J Clin Microbiol 40:556–574 [CrossRef]
     [Google Scholar]
  13. Gomez-Lopez A., Alastruey-Izquierdo A., Rodriguez D., Almirante B., Pahissa A., Rodriguez-Tudela J. L., Cuenca-Estrella M. Barcelona Candidemia Project Study Group 2008; Prevalence and susceptibility profile of Candida metapsilosis and Candida orthopsilosis : results from population-based surveillance of candidemia in Spain. Antimicrob Agents Chemother 52:1506–1509 [CrossRef]
     [Google Scholar]
  14. Iida S., Imai T., Oguri T., Okuzumi K., Yamanaka A., Moretti-Branchini L. M., Nishimura K., Mikami Y. 2005; Genetic diversity of the internal transcribed spacers (ITS) and 5.8S rRNA genes among the clinical isolates of Candida parapsilosis in Brazil and Japan. Nippon Ishinkin Gakkai Zasshi 46:133–137 [CrossRef]
     [Google Scholar]
  15. Khan Z. U., Ahmad S., Theyyathel A. M. 2008a; Detection of Aspergillus fumigatus -specific DNA, (1–3)- β -d-glucan and galactomannan in serum and bronchoalveolar lavage specimens of experimentally infected rats. Mycoses 51:129–135 [CrossRef]
     [Google Scholar]
  16. Khan Z. U., Ahmad S., Mokaddas E., Chandy R., Cano J., Cano J. 2008b; Actinomucor elegans var. kuwaitiensis isolated from the wound of a diabetic patient. Antonie van Leeuwenhoek 94:343–352 [CrossRef]
     [Google Scholar]
  17. Kocsube S., Toth M., Vagvolgyi C., Doczi I., Pesti M., Pocsi I., Szabo J., Varga J. 2007; Occurrence and genetic variability of Candida parapsilosis sensu lato in Hungary. J Med Microbiol 56:190–195 [CrossRef]
     [Google Scholar]
  18. Kurtzman C. P., Robnett C. J. 1997; Identification of clinically important ascomycetous yeasts based on nucleotide divergence in the 5′ end of the large-subunit (26S) ribosomal DNA gene. J Clin Microbiol 35:1216–1223
     [Google Scholar]
  19. Lasker B. A., Butler G., Lott T. J. 2006; Molecular genotyping of Candida parapsilosis group I clinical isolates by analysis of polymorphic microsatellite markers. J Clin Microbiol 44:750–759 [CrossRef]
     [Google Scholar]
  20. Lin D., Wu L. C., Rinaldi M. G., Lehman P. F. 1995; Three distinct genotypes within Candida parapsilosis from clinical sources. J Clin Microbiol 33:1815–1821
     [Google Scholar]
  21. Lockhart S. R., Messer S. A., Pfaller M. A., Diekema D. J. 2008a; Geographic distribution and antifungal susceptibility of the newly described species Candida orthopsilosis and Candida metapsilosis in comparison to the closely related species Candida parapsilosis . J Clin Microbiol 46:2659–2664 [CrossRef]
     [Google Scholar]
  22. Lockhart S. R., Messer S. A., Pfaller M. A., Diekema D. J. 2008b; Lodderomyces elongisporus masquerading as Candida parapsilosis as a cause of bloodstream infections. J Clin Microbiol 46:374–376 [CrossRef]
     [Google Scholar]
  23. Mokaddas E. M., Al-Sweih N. A., Khan Z. U. 2007; Species distribution and antifungal susceptibility of Candida bloodstream isolates in Kuwait: a 10-year study. J Med Microbiol 56:255–259 [CrossRef]
     [Google Scholar]
  24. Pappas P. G., Rex J. H., Lee J., Hamill R. J., Larsen R. A., Powderly W., Kauffman C. A., Hyslop N., Mangino J. E. other authors 2003; A prospective observational study of candidemia: epidemiology, therapy, and influences on mortality in hospitalized adult and pediatric patients. Clin Infect Dis 37:634–643 [CrossRef]
     [Google Scholar]
  25. Pfaller M. A., Diekema D. J. 2007; Epidemiology of invasive candidiasis: a persistent public health problem. Clin Microbiol Rev 20:133–163 [CrossRef]
     [Google Scholar]
  26. Rodriguez-Tudela J. L., Gomez-Lopez A., Alastruey-Izquierdo A., Mellado E., Bernal-Martinez L., Cuenca-Estrella M. 2007; Genotype distribution of clinical isolates of Trichosporon asahii based on sequencing of intergenic spacer 1. Diagn Microbiol Infect Dis 58:435–440 [CrossRef]
     [Google Scholar]
  27. Roy B., Meyer S. A. 1998; Confirmation of the distinct genotype groups within the form species Candida parapsilosis . J Clin Microbiol 36:216–218
     [Google Scholar]
  28. Sugita T., Nakajima M., Ikeda R., Matsushima T., Shinoda T. 2002; Sequence analysis of the ribosomal DNA intergenic spacer 1 regions of Trichosporon species. J Clin Microbiol 40:1826–1830 [CrossRef]
     [Google Scholar]
  29. Tavanti A., Davidson A. D., Gow N. A. R., Maiden M. C. J., Odds F. C. 2005; Candida orthopsilosis and Candida metapsilosis spp. nov. to replace Candida parapsilosis groups II and III. J Clin Microbiol 43:284–292 [CrossRef]
     [Google Scholar]
  30. Tavanti A., Hensgens L. A. M., Ghelardi E., Campa M., Senesi S. 2007; Genotyping of Candida orthopsilosis clinical isolates by AFLP reveals genetic diversity among independent isolates and strain maintenance within the same patient. J Clin Microbiol 45:1455–1462 [CrossRef]
     [Google Scholar]
  31. Weems J. J. Jr 1992; Candida parapsilosis : epidemiology, pathogenicity, clinical manifestations, and antimicrobial susceptibility. Clin Infect Dis 14:756–766 [CrossRef]
     [Google Scholar]
  32. Yong P. V., Chong P. P., Lau L. Y., Yeoh R. S. C., Jamal F. 2008; Molecular identification of Candida orthopsilosis isolated from blood culture. Mycopathologia 165:81–87 [CrossRef]
     [Google Scholar]
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Rapid molecular differentiation and genotypic heterogeneity among Candida parapsilosis and Candida orthopsilosis strains isolated from clinical specimens in Kuwait
J. Med. Microbiol. 58, 745 (2009); https://doi.org/10.1099/jmm.0.008235-0
/content/journal/jmm/10.1099/jmm.0.008235-0
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