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J Med Microbiol 56 (2007), 778-787; DOI: 10.1099/jmm.0.47106-0
© 2007 Society for General Microbiology
ISSN 1473-5644

Species identification and strain differentiation of clinical Candida isolates using the DiversiLab system of automated repetitive sequence-based PCR

Mark G. Wise1, Mimi Healy1, Kristy Reece1, Rebecca Smith2, Dobbie Walton1, Wendy Dutch1, Alex Renwick1, Joe Huong1, Steve Young2, Jeffrey Tarrand3 and Dimitrios P. Kontoyiannis4

1 Bacterial Barcodes Inc., Athens, GA 30602, USA

2 TriCore Reference Laboratories, Albuquerque, NM 87102, USA

3 Department of Laboratory Medicine, University of Texas M. D. Anderson Cancer Center, Houston, TX 77030, USA

4 Department of Infectious Diseases, Infection Control and Employee Health, University of Texas M. D. Anderson Cancer Center, Houston, TX 77030, USA

Correspondence
Dimitrios P. Kontoyiannis
dkontoyi{at}mdanderson.org

Received 30 November 2006
Accepted 26 February 2007

The DiversiLab system, which uses repetitive sequence-based PCR (rep-PCR) to genotype micro-organisms, was evaluated as a molecular typing tool for members of the genus Candida. Initially, 41 clinical Candida spp. (7 Candida krusei, 10 Candida parapsilosis, 7 Candida albicans, 10 Candida tropicalis and 7 Candida glabrata), previously identified at the species level by morphological and biochemical analysis, were analysed with the DiversiLab system. Species identification was confirmed by DNA sequence analysis of the contiguous internal transcribed spacer (ITS) region (ITS1–5.8S–ITS2). On the basis of an 80 % similarity threshold, rep-PCR consistently clustered like species and this set of isolates, along with five ATCC reference strains, was used to create a DNA fingerprint library with the DiversiLab software. Subsequently, an additional set of 115 clinical Candida isolates, identified biochemically as C. albicans (n=94), C. glabrata (n=8), C. parapsilosis (n=5), C. tropicalis (n=3), C. krusei (n=3) and Candida lusitaniae (n=2), isolated at a regional reference laboratory, were typed using DiversiLab. One hundred and six of these isolates clustered with members of the Candida library at >80 % similarity and thus could be assigned species identification, and initial calculations showed that identification via rep-PCR fingerprinting was 95 % concordant (101/106) with the biochemical/morphological identification. However, ITS region sequencing of the five discrepant samples, as well as the nine isolates that were <80 % similar to the database samples, showed that nine were misidentified with traditional biochemical/morphological methods. For the misidentified isolates, the sequence-based identification was in agreement with the DiversiLab clustering, yielding an actual correlation of >99 %. As traditional techniques can take several days to provide information about Candida at the genus/species level, genotyping with the DiversiLab system holds promise for more-rapid speciation of members of this genus. This system may also be useful for epidemiological studies such as source tracking that require Candida subspecies discrimination.

Abbreviations: ITS, internal transcribed spacer; rep-PCR, repetitive sequence-based PCR.




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Comparison of Typing Results Obtained for Methicillin-Resistant Staphylococcus aureus Isolates with the DiversiLab System and Pulsed-Field Gel Electrophoresis
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