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J Clin Microbiol. Nov 1995; 33(11): 2913–2919.
PMCID: PMC228606

Molecular probes for diagnosis of fungal infections.


We have developed 21 specific nucleic acid probes which target the large subunit rRNA genes from Aspergillus flavus, Aspergillus fumigatus, Aspergillus glaucus, Aspergillus niger, Aspergillus terreus, Blastomyces dermatitidis, Candida albicans, Candida (Torulopsis) glabrata, Candida guilliermondii, Candida kefyr, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, Coccidioides immitis, Cryptococcus neoformans var. gattii, Cryptococcus neoformans var. neoformans, Filobasidiella neoformans var. bacillispora, Filobasidiella neoformans var. neoformans, Histoplasma capsulatum, Pseudallescheria boydii, and Sporothrix schenckii. A section of the 28S rRNA gene from approximately 100 fungi, representing about 50 species of pathogens and commonly encountered saprophytes, was sequenced to develop universal PCR primers and species-specific oligonucleotide probes. Each step in the process of detection and identification was standardized to a common set of conditions applicable without modification to all fungi of interest and all types of clinical specimens. These steps consist of DNA extraction by boiling specimens in an alkaline guanidine-phenol-Tris reagent, amplification of a variable region of the 28S rRNA gene with universal primers, and amplicon identification by probe hybridization or DNA sequencing performed under conditions identical for all fungi. The results obtained by testing a panel of fungal isolates and a variety of clinical specimens indicate a high level of specificity.

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Selected References

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  • Arruda LK, Mann BJ, Chapman MD. Selective expression of a major allergen and cytotoxin, Asp f I, in Aspergillus fumigatus. Implications for the immunopathogenesis of Aspergillus-related diseases. J Immunol. 1992 Nov 15;149(10):3354–3359. [PubMed]
  • Buchman TG, Rossier M, Merz WG, Charache P. Detection of surgical pathogens by in vitro DNA amplification. Part I. Rapid identification of Candida albicans by in vitro amplification of a fungus-specific gene. Surgery. 1990 Aug;108(2):338–347. [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Holmes AR, Cannon RD, Shepherd MG, Jenkinson HF. Detection of Candida albicans and other yeasts in blood by PCR. J Clin Microbiol. 1994 Jan;32(1):228–231. [PMC free article] [PubMed]
  • Hopfer RL, Walden P, Setterquist S, Highsmith WE. Detection and differentiation of fungi in clinical specimens using polymerase chain reaction (PCR) amplification and restriction enzyme analysis. J Med Vet Mycol. 1993;31(1):65–75. [PubMed]
  • Kan VL. Polymerase chain reaction for the diagnosis of candidemia. J Infect Dis. 1993 Sep;168(3):779–783. [PubMed]
  • Leclerc MC, Philippe H, Guého E. Phylogeny of dermatophytes and dimorphic fungi based on large subunit ribosomal RNA sequence comparisons. J Med Vet Mycol. 1994;32(5):331–341. [PubMed]
  • Lott TJ, Kuykendall RJ, Reiss E. Nucleotide sequence analysis of the 5.8S rDNA and adjacent ITS2 region of Candida albicans and related species. Yeast. 1993 Nov;9(11):1199–1206. [PubMed]
  • Maiwald M, Kappe R, Sonntag HG. Rapid presumptive identification of medically relevant yeasts to the species level by polymerase chain reaction and restriction enzyme analysis. J Med Vet Mycol. 1994;32(2):115–122. [PubMed]
  • Makimura K, Murayama SY, Yamaguchi H. Detection of a wide range of medically important fungi by the polymerase chain reaction. J Med Microbiol. 1994 May;40(5):358–364. [PubMed]
  • Maleszka R, Clark-Walker GD. Yeasts have a four-fold variation in ribosomal DNA copy number. Yeast. 1993 Jan;9(1):53–58. [PubMed]
  • Mitchell TG, Freedman EZ, White TJ, Taylor JW. Unique oligonucleotide primers in PCR for identification of Cryptococcus neoformans. J Clin Microbiol. 1994 Jan;32(1):253–255. [PMC free article] [PubMed]
  • Nakamura H, Shibata Y, Kudo Y, Saito S, Kimura H, Tomoike H. [Detection of Aspergillus fumigatus DNA by polymerase chain reaction in the clinical samples from individuals with pulmonary aspergillosis]. Rinsho Byori. 1994 Jul;42(7):676–681. [PubMed]
  • Reddy LV, Kumar A, Kurup VP. Specific amplification of Aspergillus fumigatus DNA by polymerase chain reaction. Mol Cell Probes. 1993 Apr;7(2):121–126. [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Spreadbury C, Holden D, Aufauvre-Brown A, Bainbridge B, Cohen J. Detection of Aspergillus fumigatus by polymerase chain reaction. J Clin Microbiol. 1993 Mar;31(3):615–621. [PMC free article] [PubMed]
  • Stockman L, Clark KA, Hunt JM, Roberts GD. Evaluation of commercially available acridinium ester-labeled chemiluminescent DNA probes for culture identification of Blastomyces dermatitidis, Coccidioides immitis, Cryptococcus neoformans, and Histoplasma capsulatum. J Clin Microbiol. 1993 Apr;31(4):845–850. [PMC free article] [PubMed]

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