title: Comparison of PCR protocols for detecting Histoplasma capsulatum DNA through a multicenter study.
abstract: BACKGROUND
  A multicenter study was conducted. A panel containing DNA from Histoplasma capsulatum, as well as negative and cross-reaction controls, was sent to five different laboratories, members of the MICOMOL network from CYTED Program.
  
  
  OBJECTIVE
  The objective was to assess the accuracy of different PCR protocols to detect H. capsulatum DNA.
  
  
  METHODS
  Seven different PCR protocols were tested. They were based on PCR techniques and used unicopy and multicopy targets.
  
  
  RESULTS
  Most of these protocols (4/7) were able to detect the smallest amounts of fungal DNA (10(2)fg/μl). Overall sensitivity was 86% and specificity was 100%. The protocol based on a unicopy target (SCAR220) presented lower sensitivity (43%) but 100% specificity. The real-time protocols tested were highly reproducible, sensitive, and specific. Neither false positives nor cross-reactions were detected in any protocol.
  
  
  CONCLUSIONS
  All laboratories were able to amplify H. capsulatum DNA, and real-time PCR seems to be a promising tool to efficiently detect this pathogen in clinical samples.
authors.affiliation: Servicio de Micología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra Majadahonda-Pozuelo, Km 2, 28220 Majadahonda (Madrid), Spain. Electronic address: buitrago@isciii.es.
authors.forename: María José
authors.initials: MJ
authors.lastname: Buitrago
authors.forename: Cristina Elena
authors.initials: CE
authors.lastname: Canteros
authors.forename: Guadalupe
authors.initials: G
authors.lastname: Frías De León
authors.forename: Ángel
authors.initials: Á
authors.lastname: González
authors.forename: Manoel
authors.initials: M
authors.lastname: Marques-Evangelista De Oliveira
authors.forename: César O
authors.initials: CO
authors.lastname: Muñoz
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Ramirez
authors.forename: Adriana Isabel
authors.initials: AI
authors.lastname: Toranzo
authors.forename: Rosely
authors.initials: R
authors.lastname: Zancope-Oliveira
authors.forename: Manuel
authors.initials: M
authors.lastname: Cuenca-Estrella
mesh_terms: D004271:DNA, Fungal / Q000032:analysis*; D005656:Fungal Proteins / Q000235:genetics; D005819:Genetic Markers; D006658:Histoplasma / Q000235:genetics* / Q000302:isolation & purification; D007753:Laboratories / Q000458:organization & administration; D059021:Laboratory Proficiency Testing; D007843:Latin America; D009172:Mycology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016037:Single-Blind Method; D013030:Spain
chemical_list: D004271:DNA, Fungal; D005656:Fungal Proteins; D005819:Genetic Markers; C576552:Hcp100 protein, Histoplasma capsulatum
keywords: Diagnosis; Diagnóstico; Histoplasmosis; Multicenter; Multicéntrico; PCR