Multilocus Sequence Typing for Candida albicans Isolates from Candidemic Patients: Comparison with Southern Blot Hybridization and Pulsed-field Gel Electrophoresis Analysis

Background We evaluated the efficacy of multilocus sequence typing (MLST) for assessing the genetic relationship among Candida albicans isolates from patients with candidemia in a hospital setting. Methods A total of 45 C. albicans isolates from 21 patients with candidemia were analyzed. The MLST results were compared with results obtained by Southern blot hybridization (C1 fingerprinting) and pulsed-field gel electrophoresis (PFGE). PFGE analysis included karyotyping and restriction endonuclease analysis of genomic DNAs using BssHII (REAG-B) and SfiI (REAG-S). Results The 45 isolates yielded 20 unique diploid sequence types (DSTs) by MLST, as well as 12 karyotypes, 15 REAG-B patterns, 13 REAG-S patterns, and 14 C1 fingerprinting types. Microevolution among intra-individual isolates was detected in 6, 5, 3, 5, and 7 sets of isolates by MLST (1 or 2 allelic differences), REAG-B, REAG-S, C1 fingerprinting, and a combination of all methods, respectively. Among 20 DSTs, 17 were unique, and 3 were found in more than 1 patient. The results of 2 DSTs obtained from 9 patient isolates were in agreement with REAG and C1 fingerprinting patterns. However, the remaining DST, which was shared by 2 patient isolates, showed 2 different PFGE and C1 fingerprinting patterns. In addition, 3 sets of isolates from different patients, which differed in only 1 or 2 alleles by MLST, also exhibited different PFGE or C1 fingerprinting patterns. Conclusions MLST is highly discriminating among C. albicans isolates, but it may have some limitations in typing isolates from different patients, which may necessitate additional analysis using other techniques.

[1]  Jianping Xu,et al.  Molecular epidemiology of candidemia: evidence of clusters of smoldering nosocomial infections. , 2008, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[2]  D. Diogo,et al.  Molecular Phylogenetics of Candida albicans , 2007, Eukaryotic Cell.

[3]  A. Chowdhary,et al.  Comparison of multilocus sequence typing and Ca3 fingerprinting for molecular subtyping epidemiologically-related clinical isolates of Candida albicans. , 2006, Medical mycology.

[4]  Yee-Chun Chen,et al.  Multilocus Sequence Typing for Analyses of Clonality of Candida albicans Strains in Taiwan , 2006, Journal of Clinical Microbiology.

[5]  B. Spratt,et al.  Multilocus sequence typing of Candida albicans: strategies, data exchange and applications. , 2004, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[6]  S. Suh,et al.  Microevolution of Candida albicans Strains during Catheter-Related Candidemia , 2004, Journal of Clinical Microbiology.

[7]  C. d’Enfert,et al.  Collaborative Consensus for Optimized Multilocus Sequence Typing of Candida albicans , 2003, Journal of Clinical Microbiology.

[8]  L. Saiman,et al.  Elucidating the Origins of Nosocomial Infections with Candida albicans by DNA Fingerprinting with the Complex Probe Ca3 , 1999, Journal of Clinical Microbiology.

[9]  D. Soll,et al.  Hospital Specificity, Region Specificity, and Fluconazole Resistance of Candida albicans Bloodstream Isolates , 1998, Journal of Clinical Microbiology.

[10]  D. Soll,et al.  Colonizing populations of Candida albicans are clonal in origin but undergo microevolution through C1 fragment reorganization as demonstrated by DNA fingerprinting and C1 sequencing , 1995, Journal of clinical microbiology.

[11]  D. Stevens,et al.  Application of DNA typing methods to epidemiology and taxonomy of Candida species , 1987, Journal of clinical microbiology.