title: [Rapid identification of Histoplasma capsulatum in culture lysates].
abstract: BACKGROUND
  Histoplasma capsulatum is the agent of histoplasmosis, a deep mycosis mainly afflicting immunocompromised patients. Rapid identification of the fungus isolated from clinical specimens allows timely administration of specific treatment.
  
  
  OBJECTIVE
  To assess the ability of a dual PCR system targeting specific H. capsulatum DNA sites to identify fungal species in simple aqueous lysates from cultured fungi.
  
  
  METHODS
  We analysed the performance of two independent PCR reactions designed to amplify fragments of 111 and 279 bp included in H. capsulatum-specific gene AgM. We used 248 H. capsulatum strains and 68 isolates of other fungal species. Reaction templates consisted of aqueous lysates of cultured fungi (either in mycelial or yeast phase) obtained after three cycles of boiling and immediate cooling at 0°C. Selected strains were submitted to conventional DNA extraction and/or sequencing.
  
  
  RESULTS
  Both PCR systems performed identically. Amplification from aqueous lysates was achieved from 239 H. capsulatum strains; the remaining 9 strains only showed specific bands when purified DNA was used as template. Of all other fungal species tested, only 2 Emmonsia crescens strains amplified H. capsulatum-specific bands and sequences of their amplified PCR products matched > 97% with H. capsulatum sequences. Total test time did not exceed 7h with 96% sensitivity, 97% specificity and 99% positive predictive value.
  
  
  CONCLUSIONS
  The assay is fast, accurate and economical, and can be an alternative method for presumptive identification of H. capsulatum in simple culture lysates.
authors.affiliation: Departamento de Micología, INEI-ANLIS Dr. Carlos G. Malbrán, Buenos Aires, Argentina.
authors.forename: Belén
authors.initials: B
authors.lastname: Ibarra-Camou
authors.forename: Adriana Inés
authors.initials: AI
authors.lastname: Toranzo
authors.forename: William
authors.initials: W
authors.lastname: Lee
authors.forename: Graciela
authors.initials: G
authors.lastname: Davel
authors.forename: Cristina Elena
authors.initials: CE
authors.lastname: Canteros
mesh_terms: D001483:Base Sequence; D002912:Chrysosporium / Q000235:genetics; D003470:Culture Media; D004271:DNA, Fungal / Q000302:isolation & purification*; D005658:Fungi / Q000235:genetics / Q000254:growth & development; D006658:Histoplasma / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009172:Mycology / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D013997:Time Factors
chemical_list: D003470:Culture Media; D004271:DNA, Fungal