Two cases of relapsed HIV-associated visceral leishmaniasis successfully treated with combination therapy

Time course of parasitological and immunological parameters upon combined anti-leishmanial treatment in Case 1 (a) and Case 2 (b). Parasitemia (red circles) was measured by quantitative real-time PCR (qPCR) before and after anti-leishmanial combination (cAnti-Leish) treatment. Also shown is the CD4+？cell count in peripheral blood (empty squares; normal range: 500–1500 cells/μL). HIV RNA was？<？40 copies/mL in Case 1 and Case 2 throughout the study as tested by the Veris DxN HIV-1 assay (Beckman Coulter, Brea, CA USA). Leishmanial DNA was detected in peripheral blood by two distinct real-time (RT) PCR assays, amplifying segments of the small-subunit ribosomal RNA gene and the kinetoplast DNA (kDNA) [15]. Samples testing positive for parasitic DNA were then quantified by RT-PCR for the single copy polymerase gene (pol) of Leishmania [16]. The detection limit was 0.00005 parasite/reaction for kDNA RT-PCR and 1 parasite/reaction for pol RT-PCR. Leishmania typing was performed by hsp70 PCR [17] and L. infantum was identified in both patients. ▲: Parasitemia was quantified in bone marrow aspirates within one week before cAnti-Leish onse