Penicillium marneffei infections in immunocompromised patients are often fatal, but early treatment with appropriate antifungal agents may have a good prognosis. To select drugs for treating P. marneffei infection in Southern China, the susceptibilities of antifungal agents against P. marneffei isolates were evaluated. Twenty-five strains of P. marneffei were isolated from patients from 2008 to 2010 in Guangxi Province of China, and 14 strains were isolated from bamboo rats. The activity of voriconazole, fluconazole, itraconazole, terbinafine, and amphotericin B on the isolates was assessed by MIC analysis. Voriconazole MIC against strains of P. marneffei ranged from 0.004 to 0.25 mg/l, and it had the lowest MIC (geometric mean values, 0.04 mg/l) in comparison to the other antifungal agents. The MICs of other antifungal agents, in order of lowest to highest, were as follows: itraconazole, terbinafine, amphotericin B, and fluconazole. The results show voriconazole and itraconazole are active against P. marneffei isolates in vitro, and terbinafine may have unrealized usefulness in the treatment of infections caused by this yeast form.