title: A prospective clinical trial of a real-time polymerase chain reaction assay for the diagnosis of candidemia in nonneutropenic, critically ill adults.
abstract: BACKGROUND
  Invasive Candida infection among nonneutropenic, critically ill adults is a clinical problem that has received increasing attention in recent years. Poor performance of extant diagnostic modalities has promoted risk-based, preemptive prescribing in view of the poor outcomes associated with inadequate or delayed antifungal therapy; this risks unnecessary overtreatment. A rapid, reliable diagnostic test could have a substantial impact on therapeutic practice in this patient population.
  
  
  METHODS
  Three TaqMan-based real-time polymerase chain reaction assays were developed that are capable of detecting the main medically important Candida species, categorized according to the likelihood of fluconazole susceptibility. Assay 1 detected Candida albicans, Candida parapsilosis, Candida tropicalis, and Candida dubliniensis. Assays 2 and 3 detected Candida glabrata and Candida krusei, respectively. The clinical performance of these assays, applied to serum, was evaluated in a prospective trial of nonneutropenic adults in a single intensive care unit.
  
  
  RESULTS
  In all, 527 specimens were obtained from 157 participants. All 3 assays were run in parallel for each specimen; they could be completed within 1 working day. Of these, 23 specimens were obtained from 23 participants categorized as having proven Candida infection at the time of sampling. If a single episode of Candida famata candidemia was excluded, the estimated clinical sensitivity, specificity, and positive and negative predictive values of the assays in this trial were 90.9%, 100%, 100% and 99.8%, respectively.
  
  
  CONCLUSIONS
  These data suggest that the described assays perform well in this population for enhancing the diagnosis of candidemia. The extent to which they may affect clinical outcomes, prescribing practice, and cost-effectiveness of care remains to be ascertained.
authors.affiliation: Department of Medical Microbiology, The Royal Hospitals, Belfast, Northern Ireland. Ronan.Mcmullan@belfasttrust.hscni.net
authors.forename: R
authors.initials: R
authors.lastname: McMullan
authors.forename: L
authors.initials: L
authors.lastname: Metwally
authors.forename: P V
authors.initials: PV
authors.lastname: Coyle
authors.forename: S
authors.initials: S
authors.lastname: Hedderwick
authors.forename: B
authors.initials: B
authors.lastname: McCloskey
authors.forename: H J
authors.initials: HJ
authors.lastname: O'Neill
authors.forename: C C
authors.initials: CC
authors.lastname: Patterson
authors.forename: G
authors.initials: G
authors.lastname: Thompson
authors.forename: C H
authors.initials: CH
authors.lastname: Webb
authors.forename: R J
authors.initials: RJ
authors.lastname: Hay
mesh_terms: D000328:Adult; D002175:Candida / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002177:Candidiasis / Q000175:diagnosis* / Q000382:microbiology; D016638:Critical Illness; D017931:DNA Primers; D005260:Female; D016469:Fungemia / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012107:Research Design; D012680:Sensitivity and Specificity; D019914:Taq Polymerase
chemical_list: D017931:DNA Primers; D019914:Taq Polymerase