title: Simultaneous detection and identification of Candida, Aspergillus, and Cryptococcus species by reverse line blot hybridization.
abstract: We report on a reverse line blot (RLB) assay, utilizing fungal species-specific oligonucleotide probes to hybridize with internal transcribed spacer 2 region sequences amplified using a nested panfungal PCR. Reference and clinical strains of 16 Candida species (116 strains), Cryptococcus neoformans (five strains of Cryptococcus neoformans var. neoformans, five strains of Cryptococcus neoformans var. grubii, and six strains of Cryptococcus gatti), and five Aspergillus species (68 strains) were all correctly identified by the RLB assay. Additional fungal species (16 species and 26 strains) not represented on the assay did not exhibit cross-hybridization with the oligonucleotide probes. In simulated clinical specimens, the sensitivity of the assay for Candida spp. and Aspergillus spp. was 10(0.5) cells/ml and 10(2) conidia/ml, respectively. This assay allows sensitive and specific simultaneous detection and identification of a broad range of fungal pathogens.
authors.affiliation: Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Research, Westmead Hospital, New South Wales, Australia. geoffrey_playford@health.qld.gov.au
authors.forename: E Geoffrey
authors.initials: EG
authors.lastname: Playford
authors.forename: Fanrong
authors.initials: F
authors.lastname: Kong
authors.forename: Ying
authors.initials: Y
authors.lastname: Sun
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Catriona
authors.initials: C
authors.lastname: Halliday
authors.forename: Tania C
authors.initials: TC
authors.lastname: Sorrell
mesh_terms: D001230:Aspergillus / Q000235:genetics* / Q000302:isolation & purification*; D001483:Base Sequence; D002175:Candida / Q000235:genetics* / Q000302:isolation & purification*; D003455:Cryptococcus neoformans / Q000235:genetics* / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000235:genetics; D006801:Humans; D015336:Molecular Probe Techniques / Q000706:statistics & numerical data; D009172:Mycology / Q000379:methods* / Q000706:statistics & numerical data; D015345:Oligonucleotide Probes; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D004271:DNA, Fungal; D015345:Oligonucleotide Probes