title: Clinical validation of a multiplex real-time PCR assay for detection of invasive candidiasis in intensive care unit patients.
abstract: BACKGROUND
  New techniques, such as those based on multiplex quantitative real-time PCR (MRT-PCR), can improve the detection of invasive candidiasis (IC).
  
  
  METHODS
  We prospectively studied 63 intensive care unit patients with suspected IC and 40 healthy controls. Blood cultures and MRT-PCR were performed at day 0 and +2, +7, +14 and +21 days in all patients. In addition, β-d-glucan (BDG) and Candida albicans germ tube antibody (CAGTA) were quantified.
  
  
  RESULTS
  IC was confirmed in 27 patients. Colonization was significantly higher in patients with IC (96% versus 64%, P = 0.002). The sensitivity, specificity, positive predictive value and negative predictive value of MRT-PCR for the diagnosis of IC were 96.3%, 97.3%, 92.8% and 98.7%, respectively. The positive predictive value and specificity were significantly higher for MRT-PCR than for BDG and CATGA. MRT-PCR performed very well, especially in deep-seated IC (sensitivity 90.9% versus 45.4% for blood culture; P = 0.06). As regards the most appropriate clinical sample for DNA amplification, in this study whole blood and serum presented similar results.
  
  
  CONCLUSIONS
  MRT-PCR appears to be a useful test for confirming a diagnosis of IC in critically ill patients, especially in those with deep-seated disease. Its high sensitivity and positive predictive value make it a much more efficient tool for the management of IC than other diagnostic procedures and clinical scores.
authors: Servicio de Enfermedades Infecciosas, Hospital Ramón y Cajal, Instituto Ramón y Cajal de Investigaciones Sanitarias, Madrid, Spain fortunabete@gmail.com.
authors: J
authors: J
authors: Fortún
authors: Servicio de Enfermedades Infecciosas, Hospital Ramón y Cajal, Instituto Ramón y Cajal de Investigaciones Sanitarias, Madrid, Spain.
authors: Y
authors: Y
authors: Meije
authors: Servicio de Micología, Centro Nacional de Microbiología, Majadahonda, Spain.
authors: M J
authors: MJ
authors: Buitrago
authors: Servicio de Micología, Centro Nacional de Microbiología, Majadahonda, Spain.
authors: S
authors: S
authors: Gago
authors: Servicio de Micología, Centro Nacional de Microbiología, Majadahonda, Spain.
authors: L
authors: L
authors: Bernal-Martinez
authors: Servicio de Microbiología, Hospital Universitario y Politécnico La Fe, Valencia, Spain.
authors: J
authors: J
authors: Pemán
authors: Fontlab2000, Barcelona, Spain.
authors: M
authors: M
authors: Pérez
authors: Servicio de Microbiología, Hospital Ramón y Cajal, Instituto Ramón y Cajal de Investigaciones Sanitarias, Madrid, Spain.
authors: E
authors: E
authors: Gómez-G Pedrosa
authors: Servicio de Enfermedades Infecciosas, Hospital Ramón y Cajal, Instituto Ramón y Cajal de Investigaciones Sanitarias, Madrid, Spain.
authors: N
authors: N
authors: Madrid
authors: Servicio de Enfermedades Infecciosas, Hospital Ramón y Cajal, Instituto Ramón y Cajal de Investigaciones Sanitarias, Madrid, Spain.
authors: V
authors: V
authors: Pintado
authors: Servicio de Enfermedades Infecciosas, Hospital Ramón y Cajal, Instituto Ramón y Cajal de Investigaciones Sanitarias, Madrid, Spain.
authors: P
authors: P
authors: Martín-Dávila
authors: Servicio de Enfermedades Infecciosas, Hospital Ramón y Cajal, Instituto Ramón y Cajal de Investigaciones Sanitarias, Madrid, Spain.
authors: J
authors: J
authors: Cobo
authors: Servicio de Enfermedades Infecciosas, Hospital Ramón y Cajal, Instituto Ramón y Cajal de Investigaciones Sanitarias, Madrid, Spain.
authors: G
authors: G
authors: Fresco
authors: Servicio de Enfermedades Infecciosas, Hospital Ramón y Cajal, Instituto Ramón y Cajal de Investigaciones Sanitarias, Madrid, Spain.
authors: S
authors: S
authors: Moreno
authors: Servicio de Micología, Centro Nacional de Microbiología, Majadahonda, Spain.
authors: M
authors: M
authors: Cuenca-Estrella
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D058365:Candidiasis, Invasive / Q000097:blood* / Q000175:diagnosis*; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D007362:Intensive Care Units / Q000592:standards*; D008297:Male; D008875:Middle Aged; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D055815:Young Adult
keywords: CAGTA; Candida; PCR; β-d-glucan