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The standard treatment of amphotericin B (AmB) induction followed by itraconazole consolidation still results in 15% to 30% mortality. This study aimed to investigate the potential of AmB and flucytosine (5FC) combination therapy to enhance antifungal activity.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      The\n                      <jats:italic>in vitro<\/jats:italic>\n                      antifungal activity of AmB and 5FC alone and in combination against 60\n                      <jats:italic>T. marneffei<\/jats:italic>\n                      clinical isolates was evaluated using a validated colorimetric antifungal susceptibility assay and the checkerboard method. The minimum inhibitory concentration (MIC) was defined as the lowest drug concentration inhibiting\u2009\u2265\u200995% fungal growth (MIC\n                      <jats:sub>95<\/jats:sub>\n                      ) for both AmB and 5FC. The combination effect between AmB and 5FC against\n                      <jats:italic>T. marneffei<\/jats:italic>\n                      was determined using fractional inhibitory concentration index. Combination effects were further tested using a time-kill assay.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The MIC\n                      <jats:sub>95<\/jats:sub>\n                      was 0.25 \u2013 2 \u03bcg\/mL (geometric mean [GM] 0.68 \u03bcg\/mL) for AmB, and 0.03 \u2013 0.5 \u03bcg\/mL (GM 0.28 \u03bcg\/mL) for 5FC. Full synergy was observed in 4 isolates (7%), and indifference was observed in the remaining 56 isolates (93%). The time-kill experiments revealed a concentration-dependent fungicidal activity of AmB, and concentration-independent fungistatic effect of 5FC. Synergy between AmB and 5FC was confirmed, showing greater than 2-log\n                      <jats:sub>10<\/jats:sub>\n                      reduction in colony forming units when used in combination. No antagonism was observed.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      Our study demonstrated\n                      <jats:italic>in vitro<\/jats:italic>\n                      evidence of synergistic activity between AmB and 5FC against\n                      <jats:italic>T. marneffei<\/jats:italic>\n                      , providing the evidence to support\n                      <jats:italic>in vivo<\/jats:italic>\n                      and clinical trial testing of AmB and 5FC combination therapy, and dosing reduction strategies of 5FC.\n                    <\/jats:p>\n                  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