AI Article Synopsis
- * Seven PCR techniques were tested, with four demonstrating the ability to identify very low amounts of fungal DNA, achieving an overall sensitivity of 86% and a specificity of 100%.
- * Results indicated that while all labs successfully amplified the DNA, real-time PCR protocols showed high reliability and potential for clinical use with no false positives or cross-reactions noted.
Article 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.
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| http://dx.doi.org/10.1016/j.riam.2013.03.004 | DOI Listing |
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