Abstract
Fungal burden in the cerebrospinal fluid is an important determinant of mortality in cryptococcal meningitis, but its use in aiding clinical decision making is hampered by the time involved to perform quantitative cultures. Here, we demonstrate the potential of flow cytometry as a novel and rapid technique to address this issue.
Highlights
Cryptococcal meningitis (CM) remains one of the commonest causes of meningitis in sub-Saharan Africa and a significant cause of death among persons with HIV-1 infection (1, 2)
cerebrospinal fluid (CSF) fungal burden was assessed with quantitative culture (5) and cryptococcal antigen (CrAg) titer (CrAg lateral flow assay [LFA]; Immy, USA), as previously described (6)
The volume of remaining CSF was measured and the cells pelleted using centrifugation; this was incubated on ice with an amine viability dye (Aqua; Invitrogen) and anti-CD45-PECy5.5 (BioLegend) and at room temperature with flow-activated cell sorting (FACS) lysing solution (BD Biosciences), protected from light at all times
Summary
Cryptococcal meningitis (CM) remains one of the commonest causes of meningitis in sub-Saharan Africa and a significant cause of death among persons with HIV-1 infection (1, 2). CSF fungal burden was assessed with quantitative culture (5) and cryptococcal antigen (CrAg) titer (CrAg lateral flow assay [LFA]; Immy, USA), as previously described (6). Sixty HIV-infected patients with cryptococcal meningitis were enrolled, with a median CD4 count of 34 cells/l of CSF.
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