Molecular probes for diagnosis of fungal infections

G. S. Sandhu, B. C. Kline, L. Stockman, G. D. Roberts

Research output: Contribution to journalArticle

321 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)2913-2919
Number of pages7
JournalJournal of Clinical Microbiology
Volume33
Issue number11
StatePublished - 1995

Fingerprint

Molecular Probes
Mycoses
Candida
rRNA Genes
Candida glabrata
Fungi
Aspergillus
Cryptococcus gattii
Pseudallescheria
Blastomyces
Nucleic Acid Probes
Coccidioides
Sporothrix
Candida tropicalis
Histoplasma
Aspergillus flavus
Cryptococcus neoformans
Aspergillus fumigatus
Oligonucleotide Probes
Aspergillus niger

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Sandhu, G. S., Kline, B. C., Stockman, L., & Roberts, G. D. (1995). Molecular probes for diagnosis of fungal infections. Journal of Clinical Microbiology, 33(11), 2913-2919.

Molecular probes for diagnosis of fungal infections. / Sandhu, G. S.; Kline, B. C.; Stockman, L.; Roberts, G. D.

In: Journal of Clinical Microbiology, Vol. 33, No. 11, 1995, p. 2913-2919.

Research output: Contribution to journalArticle

Sandhu, GS, Kline, BC, Stockman, L & Roberts, GD 1995, 'Molecular probes for diagnosis of fungal infections', Journal of Clinical Microbiology, vol. 33, no. 11, pp. 2913-2919.
Sandhu GS, Kline BC, Stockman L, Roberts GD. Molecular probes for diagnosis of fungal infections. Journal of Clinical Microbiology. 1995;33(11):2913-2919.
Sandhu, G. S. ; Kline, B. C. ; Stockman, L. ; Roberts, G. D. / Molecular probes for diagnosis of fungal infections. In: Journal of Clinical Microbiology. 1995 ; Vol. 33, No. 11. pp. 2913-2919.
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