Infections caused by yeast of the genus Candida are the most common fungal infections, being Candida albicans the most common isolated species among them. The rapid identification of this yeast is mostly based on the production of germ tube in human or animal serum. This study describes the use of 12 different liquid media for germ tube production at 2, 2.5, 3 and 4 h. We examined 193 yeasts, including 157 (81.3%) C. albicans and 36 (18.7%) Candida tropicalis for the production of germ tube. The germ tube production of C. albicans was mostly observed in human serum (98%) followed by rabbit serum (89.8%), brain heart infusion broth (84%) and sheep serum (74.5%) at 2 h. An incubation time exceeding 2 h i.e. 2.5 h or later, C. tropicalis strains were observed to produce germ tubes. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for germ tube production of human serum at 2 h were 98%, 100%, 100% and 92.3% respectively. In all tested sera, an incubation period of more than 2 h improves the sensitivity, but decreases the specificity as well as PPV and NPV of germ tube test (GTT). In conclusion, human serum was observed to be the most appropriate medium to be preferred for GTT, with an incubation period of 2 h.