Asymptomatic cryptococcal antigen prevalence detected by lateral flow assay in hospitalised HIV-infected patients in São Paulo, Brazil.

OBJECTIVE: To determine the prevalence of asymptomatic cryptococcal antigen (CRAG) using lateral flow assay (LFA) in hospitalised HIV-infected patients with CD4 counts <200 cells/µl. METHODS: Hospitalised HIV-infected patients were prospectively recruited at Instituto de Infectologia Emilio Ribas, a tertiary referral hospital to HIV-infected patients serving the São Paulo State, Brazil. All patients were >18 years old without prior cryptococcal meningitis, without clinical suspicion of cryptococcal meningitis, regardless of antiretroviral (ART) status, and with CD4 counts <200 cells/µl. Serum CRAG was tested by LFA in all patients, and whole blood CRAG was tested by LFA in positive cases. RESULTS: We enrolled 163 participants of whom 61% were men. The duration of HIV diagnosis was a median of 8 (range, 1-29) years. 26% were antiretroviral (ART)-naïve, and 74% were ART-experienced. The median CD4 cell count was 25 (range, 1-192) cells/µl. Five patients (3.1%; 95%CI, 1.0-7.0%) were asymptomatic CRAG-positive. Positive results cases were cross-verified by performing LFA in whole blood. CONCLUSIONS: 3.1% of HIV-infected inpatients with CD4 <200 cells/µl without symptomatic meningitis had cryptococcal antigenemia in São Paulo, suggesting that routine CRAG screening may be beneficial in similar settings in South America. Our study reveals another targeted population for CRAG screening: hospitalised HIV-infected patients with CD4 <200 cells/µl, regardless of ART status. Whole blood CRAG LFA screening seems to be a simple strategy to prevention of symptomatic meningitis.

Disseminated amphotericin-resistant fusariosis in acute leukemia patients: report of two cases.

Disseminated fusariosis has emerged as a significant, usually fatal infection in immunocompromised hosts despite antifungal treatment. We describe here two patients with acute leukemia who developed disseminated amphotericin-resistant fusariosis, and review of six studies of cases series in the literature. Two Fusarium solani strains were isolated from blood and skin cultures of one patient, and one strain from the blood culture of the second patient. Both patients died despite antifungal treatment. Strains were identified by sequencing of ITS1 and ITS4 regions. Random amplified polymorphic DNA analysis of the three F. solani isolates showed a low degree of similarity. Screening for Fusarium spp. contaminants within our facility was negative. Using the CLSI M-38-A2 broth dilution method and E tests(®), we found that the MICs were low for voriconazole (0.12 and 0.5 mg/L, respectively), unexpectedly high for amphotericin B (≥8 and ≥32 µg/mL, respectively) and itraconazole (≥16 mg/ml). Patients with leukemia or persistent neutropenia should be assessed for disseminated fungal infections, including biopsy and skin cultures. Antifungal susceptibility tests are important due to the possibility of the strains being amphotericin resistant. Treatments must be aggressive, with high doses of antifungals or combined therapy.

Cerebral infection caused by Cryptococcus gattii: a case report and antifungal susceptibility testing.

We report a clinical case of cerebral infection caused by Cryptococcus gattii in a 10 year-old boy. Clinical and laboratory exams did not demonstrate any apparent immunosuppressed state (HIV antibody and the tuberculin skin tests, both negative, were performed; blood cells count and immunoglobulin levels were within normality). Treatment was begun with amphotericin B-deoxycholate but renal toxicity signs led to its substitution by fluconazole. The infection proceeded even after treatment with fluconazole. In vitro determination of minimum inhibitory concentration values were high for itraconazole (= or > 2 microg/ml), fluconazole and 5-flucytosine (= or > 64 microg/ml) and low for amphotericin B (1.0 microg/ml). Renal toxicity signs, induced by amphotericin B, progression of infection after fluconazole, and likely in vivo resistance to this triazole made this case difficult to treat. In vitro drug interaction tests confirmed probable synergism between amphotericin B and 5-flucytosine (frational inhibitory concentration - FIC = 0.375). In contrast, a probable additive effect was observed for amphotericin B and fluconazole (FIC = 0.75). Initial treatment of persistent high intracranial pressure was insufficient and neurological surgery was necessary. Antifungal susceptibility tests and Cryptococcus species identification were important in selecting appropriate antifungal therapy.

Antifungal drug susceptibility profile of Pichia anomala isolates from patients presenting with nosocomial fungemia.

In vitro susceptibility of 58 isolates of Pichia anomala to five antifungal drugs using two broth microdilution methods (CLSI and EUCAST) was analyzed. Low susceptibility to itraconazole was observed. Fluconazole, voriconazole, amphotericin B, and caspofungin showed good antifungal activity, although relatively high drug concentrations were necessary to inhibit the isolates.