title: Acquired Multidrug Antifungal Resistance in Candida lusitaniae during Therapy.
abstract: Candida lusitaniae is usually susceptible to echinocandins. Beta-1,3-glucan synthase encoded by FKS genes is the target of echinocandins. A few missense mutations in the C. lusitaniae FKS1 hot spot 1 (HS1) have been reported. We report here the rapid emergence of antifungal resistance in C. lusitaniae isolated during therapy with amphotericin B (AMB), caspofungin (CAS), and azoles for treatment of persistent candidemia in an immunocompromised child with severe enterocolitis and visceral adenoviral disease. As documented from restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analysis, the five C. lusitaniae isolates examined were related to each other. From antifungal susceptibility and molecular analyses, 5 different profiles (P) were obtained. These profiles included the following: profile 1 (P1) (CAS MIC [μg/ml], 0.5; fluconazole [FLC] MIC, 0.25), determined while the patient was being treated with liposomal AMB for 3 months; P2 (FLC MIC [μg/ml], 0.25; CAS MIC, 4), while the patient was being treated with CAS for 2 weeks; P3 (CAS MIC [μg/ml], 0.5; FLC MIC, 32), while the patient was being treated with azoles and CAS initially followed by azoles alone for a week; P4 (CAS MIC [μg/ml], 8; FLC MIC, 8), while the patient was being treated with both drugs for 3 weeks; and P5 (AMB MIC [μg/ml], 0.125; CAS MIC, 8), while the patient was being treated with AMB and FLC for 2 weeks. CAS resistance was associated with resistance not only to micafungin and anidulafungin but also to AMB. Analysis of CAS resistance revealed 3 novel FKS1 mutations in CAS-resistant isolates (S638Y in P2; S631Y in P4; S638P in P5). While S638Y and -P are within HS1, S631Y is in close proximity to this domain but was confirmed to confer candin resistance using a site-directed mutagenesis approach. FLC resistance could be linked with overexpression of major facilitator gene 7 (MFS7) in C. lusitaniae P2 and P4 and was associated with resistance to 5-flurocytosine. This clinical report describes resistance of C. lusitaniae to all common antifungals. While candins or azole resistance followed monotherapy, multidrug antifungal resistance emerged during combined therapy.
authors.affiliation: Pediatric Infectious Diseases and Vaccinology Unit, Department of Paediatrics, University Hospital Center, Lausanne, Switzerland Service of Infectious Diseases, Department of Internal Medicine, University Hospital Center, Lausanne, Switzerland.
authors.forename: Sandra A
authors.initials: SA
authors.lastname: Asner
authors.affiliation: Service of Infectious Diseases, Department of Internal Medicine, University Hospital Center, Lausanne, Switzerland.
authors.forename: Stefano
authors.initials: S
authors.lastname: Giulieri
authors.affiliation: Pediatric Haematology and Oncology Unit, Department of Pediatrics, University Hospital Center Lausanne, Switzerland.
authors.forename: Manuel
authors.initials: M
authors.lastname: Diezi
authors.affiliation: Service of Infectious Diseases, Department of Internal Medicine, University Hospital Center, Lausanne, Switzerland.
authors.forename: Oscar
authors.initials: O
authors.lastname: Marchetti
authors.affiliation: Institute of Microbiology, Department of Laboratory, University of Lausanne and University Hospital Center, Lausanne, Switzerland dominique.sanglard@chuv.ch.
authors.forename: Dominique
authors.identifier: http://orcid.org/0000-0002-5244-4178
authors.initials: D
authors.lastname: Sanglard
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000627:therapeutic use*; D002175:Candida / Q000187:drug effects*; D002177:Candidiasis / Q000188:drug therapy* / Q000382:microbiology*; D004271:DNA, Fungal / Q000235:genetics; D016903:Drug Monitoring; D026141:Drug Resistance, Multiple, Fungal / Q000235:genetics; D005260:Female; D005690:Galactose / Q000031:analogs & derivatives; D006801:Humans; D016867:Immunocompromised Host; D007223:Infant; D015470:Leukemia, Myeloid, Acute / Q000150:complications / Q000188:drug therapy; D008351:Mannans / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D012150:Polymorphism, Restriction Fragment Length; D047071:beta-Glucans / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D004271:DNA, Fungal; D008351:Mannans; D047071:beta-Glucans; C012990:galactomannan; D005690:Galactose