title: Isolation and drug susceptibility of Candida parapsilosis sensu lato and other species of C. parapsilosis complex from patients with blood stream infections and proposal of a novel LAMP identification method for the species.
abstract: Candida parapsilosis complex (CPC) is the third Candida species isolated in blood cultures of patients from our Hospital, following C. albicans and C. tropicalis. From 2006 to 2010, the median annual distribution of CPC was 8 cases/year. Records of 36 patients were reviewed. CPC were 31 (86.1%) C. parapsilosis; 4 (11.1%) C. orthopsilosis; and 1 (2.8%) C. metapsilosis. Clinical characteristics were central venous catheter, 34 (94.4%); parental nutrition, 25 (70%); surgery, 27 (57.9%); prior bacteremia, 20 (51.3%); malignancy, 18 (50%). General mortality was 47.2%. Death was higher in immunosuppressed patients (17 vs. 11; p = 0.003). Three out four (75%) patients with C. orthopsilosis and 14 out 31 (45.2%) with C. parapsilosis died (p = 0.558). Thirty-nine individual isolates were tested for susceptibility to seven antifungal drugs, with MICs values showing susceptibility to all of them. Two isolates, one C. orthopsilosis and one C. parapsilosis, had fluconazole MIC = 4 μg/mL. Differentiation among CPC has implication in caring for patients with invasive candidiasis since there are differences in virulence, pathogenicity and drug susceptibility. A method targeting the topoisomerase II gene based on loop-mediated isothermal amplification (LAMP) was developed. LAMP emerges as a promising tool for the identification of fungal species due to the high sensitivity and specificity. LAMP can be performed at the point-of-care, being no necessary the use of expensive equipment. In our study, the method was successful comparing to the DNA sequencing and proved to be a reliable and fast assay to distinguish the three species of CPC.
authors.affiliation: Infectious Diseases Division, School of Medical Sciences, University of Campinas, Rua Tessália Vieira de Carvalho, 126 Cidade Universitária Zeferino Vaz, Campinas, Sao Paulo, 13083-887, Brazil, trabasso@fcm.unicamp.br.
authors.forename: Plinio
authors.initials: P
authors.lastname: Trabasso
authors.forename: Tetsuhiro
authors.initials: T
authors.lastname: Matsuzawa
authors.forename: Renata
authors.initials: R
authors.lastname: Fagnani
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Muraosa
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Tominaga
authors.forename: Mariangela Ribeiro
authors.initials: MR
authors.lastname: Resende
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Kamei
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Mikami
authors.forename: Angelica Zaninelli
authors.initials: AZ
authors.lastname: Schreiber
authors.forename: Maria Luiza
authors.initials: ML
authors.lastname: Moretti
mesh_terms: D000935:Antifungal Agents / Q000627:therapeutic use; D001483:Base Sequence; D002175:Candida / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D058387:Candidemia / Q000175:diagnosis* / Q000188:drug therapy / Q000382:microbiology* / Q000401:mortality; D002177:Candidiasis / Q000175:diagnosis* / Q000188:drug therapy / Q000382:microbiology* / Q000401:mortality; D004271:DNA, Fungal / Q000235:genetics; D025141:Drug Resistance, Fungal; D005260:Female; D015725:Fluconazole / Q000627:therapeutic use; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA
chemical_list: D000935:Antifungal Agents; D004271:DNA, Fungal; D015725:Fluconazole