Phaeoacremonium parasiticum is an agent of opportunistic phaeohyphomycosis belonging to a genus encompassing numerous recently described and soon-to-be-described, difficult-to-identify human pathogens. It appears in the literature to be an uncommon etiologic agent, yet we encountered several cases in a single year. Each presented problems in laboratory identification and case management. We present two cases of invasive disease with definite identification and susceptibility results. These cases are analyzed in relation to a brief review of previous cases known to have been caused by this species. Our first case involved a 40-year-old male cardiac transplant recipient with multiple localized skin lesions. The second featured a 31-year-old female with aplastic anemia and prolonged neutropenia who developed disseminated disease, with multiple positive blood cultures and skin lesions. Both patients died despite aggressive surgical and antifungal therapy. Fungal susceptibility testing showed that our isolates appeared to be susceptible to amphotericin B, itraconazole, voriconazole, ravuconazole, and posaconazole. Because phenotypic identification of Phaeoacremonium is notably problematic, sequence-based confirmation was performed using a recently proposed standard based on use of a segment of the 5′ end of the β-tubulin gene. Sequences from both isolates involved in the cases were over 99% similar to the corresponding sequence of the ex-type isolate of P. parasiticum. The close DNA similarity, corroborated by relevant morphological similarities (e.g., long, thin phialides and tuberculate hyphae bearing warts up to 3 μm high), confirms these two isolates as P. parasiticum. Copyright © 2006, American Society for Microbiology. All Rights Reserved.