1. Comparison of PCR protocols for detecting Histoplasma capsulatum DNA through a multicenter study.
- Author
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Buitrago MJ, Canteros CE, Frías De León G, González Á, Marques-Evangelista De Oliveira M, Muñoz CO, Ramirez JA, Toranzo AI, Zancope-Oliveira R, and Cuenca-Estrella M
- Subjects
- Fungal Proteins genetics, Genetic Markers, Histoplasma isolation & purification, Laboratories organization & administration, Laboratory Proficiency Testing, Latin America, Real-Time Polymerase Chain Reaction methods, Reproducibility of Results, Sensitivity and Specificity, Single-Blind Method, Spain, DNA, Fungal analysis, Histoplasma genetics, Mycology methods, Polymerase Chain Reaction methods
- 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., Aims: 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., (Copyright © 2012 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.)
- Published
- 2013
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