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Journal of Clinical Microbiology, March 2006, p. 876-880, Vol. 44, No. 3
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.3.876-880.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Simultaneous Detection and Identification of Candida, Aspergillus, and Cryptococcus Species by Reverse Line Blot Hybridization

E. Geoffrey Playford,1,2,3* Fanrong Kong,1,2 Ying Sun,1,4 Hui Wang,1,5 Catriona Halliday,1 and Tania C. Sorrell1,2

Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Research, Westmead Hospital, Westmead, New South Wales 2145, Australia,1 Department of Medicine, University of Sydney, Sydney, Australia,2 Princess Alexandra Hospital, Brisbane, Australia,3 Public Health College, Tianjin Medical University, Tianjin, People's Republic of China,4 Department of Dermatology, Wuhan First Hospital, Wuhan, Hubei Province, People's Republic of China5

Received 7 July 2005/ Returned for modification 15 September 2005/ Accepted 24 October 2005

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 100.5 cells/ml and 102 conidia/ml, respectively. This assay allows sensitive and specific simultaneous detection and identification of a broad range of fungal pathogens.

* Corresponding author. Mailing address: Infection Management Services, Princess Alexandra Hospital, Ipswich Rd., Woolloongabba, Q 4102, Australia. Phone: 61-7-3240 2329. Fax: 61-7-3240 5540. E-mail: geoffrey_playford{at}health.qld.gov.au.

Journal of Clinical Microbiology, March 2006, p. 876-880, Vol. 44, No. 3
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.3.876-880.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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