We describe a case of Pichia farinosa bloodstream infection in a lymphoma patient. Phenotypic methods failed to identify the isolate, which was identified by sequence-based methods. This case highlights the importance of implementing molecular methods for the identification of rare fungal pathogens.
CASE REPORTA 13-year-old boy with anaplastic large-cell lymphoma presented with fever for several hours and vomiting. The diagnosis of lymphoma was made 3 months previously, and a Broviac catheter was inserted into the right jugular vein. The patient was treated with chemotherapy that included systemic dexamethasone, cyclophosphamide, and methotrexate, along with prophylactic antimicrobial therapy with trimethoprim-sulfamethoxazole. The patient had one episode of Pseudomonas sp. sepsis that was treated with ciprofloxacin. The fifth chemotherapy course was administered 2 weeks prior to his admission, and severe neutropenia (absolute neutrophil count, Ͻ500 cells/ l) was present from 2 days before his admission. On admission, the patient appeared alert, his temperature was 40°C, his heart rate was 160 beats per minute, and the rest of the physical examination was normal. Treatment with piperacillin and gentamicin was initiated after drawing of two sets of blood cultures (BACTEC; Becton Dickinson) through the Broviac catheter. Three days after his admission, growth of yeasts was detected microscopically in three blood culture sets, and local redness and swelling were observed along the catheter tunnel. Treatment with amphotericin B was started, and the Broviac catheter was removed.The fungus that grew on Sabouraud dextrose agar medium was identified by using the API 20 C AUX identification system (bioMérieux, France). The system code (6412044) was interpreted as Candida boidinii (% identification ϭ 99.8%; T ϭ 0.51). Further characterization was attempted by using the ID 32 C identification system (bioMérieux, France). The system code (1001711217) with the closest match was Pichia farinosa, but since two tests contradicted with the software's data (assimilation in L-sorbose and lack of assimilation in N-acetylglucosamine), it was reported as an "unacceptable profile." Molecular identification was attempted by amplification and sequencing of a fragment that includes internal transcribed spacer 1 (ITS 1), the 5.8S rRNA gene, and ITS 2 (9) as previously described, with minor modifications of the PCR conditions. The PCR assay was performed with 1 l DNA in a total volume of 50 l. The assay mixture contained 5 l BIOTAQ 10ϫ PCR buffer (Bioline, United Kingdom), 3.0 mM magnesium chloride, 1.0 l of a 10 mM deoxynucleoside triphosphate mixture, 50 pmol of each of the respective primers, and 2.5 U BIOTAQ DNA polymerase (Bioline, United Kingdom) per 50 l. After initial denaturation at 94°C for 5 min, 35 cycles were performed, consisting of a denaturation step at 94°C for 40 s, an annealing step at 49°C for 40 s, and an extension step at 72°C for 45 s, with a final extension step at 72°C for 2 min following the last cycle. For amplifi...