Candida albicans is the most common fungal pathogen. Galleria mellonella is widely used as an infection model host. Nevertheless, the G. mellonella–C. albicans infection model had not been optimized for drug evaluation before this study. In this work, we revealed that 5×105 colony forming unit (CFU)/larva was a suitable inoculum to optimize the G. mellonella–C. albicans infection model in order to evaluate antifungal agents. Using our optimized model, the antifungal effect of fluconazole, amphotericin B and flucytosine, and the synergy between amphotericin B and flucytosine were successfully verified. Thus, this study provides a rapid, inexpensive and reliable way to evaluate antifungals in vivo .