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Updated: May 12, 2026
Optimized PCR-based Detection of Mycoplasma
Published on: June 20, 2011
Comparison of PCR protocols for detecting Histoplasma capsulatum DNA through a multicenter study
María José Buitrago1, Cristina Elena Canteros, Guadalupe Frías De León
1Servicio de Micología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra Majadahonda-Pozuelo, Km 2, 28220 Majadahonda (Madrid), Spain.
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.
