The aim of this study was to screen for C. dubliniensis using the germ tube test with human pooled serum (HPS) in clinical isolates and investigate the prevalence of C. dubliniensis in Korea. Among 1,854 yeast strains isolated, 1,404 strains of C. albicans (on the basis of positive results of the germ tube test) and 192 germ tube-negative yeast strains were examined. All 1,596 clinical isolates were examined using the germ tube test with HPS, the differential temperature, and NaCl tolerance test. Only 81 isolates that did not grow at 45°C nor on Sabouraud 6.5% NaCl broth were selected and tested using the VITEK 2 ID-YSTsystem and the multiplex-PCR assay for the study. The two strains, C. dubliniensis ATCC MYA-646 and KCTC 17427 failed to produce germ tubes in HPS but produced them in fresh rabbit serum (FRS) and fetal bovine serum (FBS). No C. dubliniensis was found in this study population. The results of this study suggest that the germ tube test with HPS in combination with FRS or FBS can be used for discriminating between C. albicans and C. dubliniensis strains and that the prevalence of C. dubliniensis appears to be extremely low in Korea. J. Clin. Lab. Anal. 24:145–148, 2010. © 2010 Wiley-Liss, Inc.