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Original Articles
Molecular Typing of Urinary Isolates of Candida tropicalis by Pulsed-Field Gel Electrophoresis (PFGE) and Random Amplified Polymorphic DNA (RAPD) Analysis
Joon Rho, M.D., Chul Sung Kim, M.D., Sook Jin Jang, M.D.*, Jong Hee Shin, M.D.† Soon Pal Suh, M.D.† and Dong Wook Ryang, M.D.†
Departments of Urology and Clinical Pathology*, Chosun University College of Medicine, Department of Clinical Pathology†, Chonnam National University Medical School, Gwangju, Korea
Vol.33 Num.6 (p392~403)
Background:Despite the recognized increase of frequency of candiduria due to Candida tropicalis, little was known of its molecular epidemiology. We applied PFGE and RAPD assay for urinary C. tropicalis isolates and evaluated the utilities of PFGE and RAPD for the epidemiological typing of C. tropicalis isolates.
Methods:A total of urinary 57 isolates of C. tropicalis from 40 patients at two hospitals was analyzed. PFGE analysis were performed by electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG) using two restriction enzymes (BssHII and SfiI). For RAPD, a total of 31 primers (30 random 10-mer primers and M13 primer) were used.
Results:EK and RAPD analysis showed the same or similar patterns among the isolates. REAG with BssHII separated 57 isolates into 28 distinct types. Six patterns were generated by REAG with using SfiI. By combining the two REAG, a total of 31 different DNA types were identified among 57 isolates from 40 patients. Three strain types were common to 23 isolates from 12 patients of a University Hospital, which suggested possible nosocomial transmission. In 19 patients with serial urinary isolates, the sequential strains from each patient exhibited the same REAG pattern.
Conclusion:These suggest that REAG with BssHII and SfiI is useful for the investigation of molecular epidemiology of C. tropicalis isolates. In addition, some clusters of C. tropicalis isolates with the same DNA type suggest that nosocomial transmission may occur.
Keywords : Candida tropicalis, Pulsed-field gel electrophoresis (PFGE), Random amplified polymorphic DNA (RAPD), Candiduria