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.
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.
Authors: María Guadalupe Frías-De-León; José Antonio Ramírez-Bárcenas; Gabriela Rodríguez-Arellanes; Oscar Velasco-Castrejón; Maria Lucia Taylor; María Del Rocío Reyes-Montes Journal: Folia Microbiol (Praha) Date: 2016-10-10 Impact factor: 2.099
Authors: Sara Gago; Cristina Esteban; Clara Valero; Oscar Zaragoza; Jorge Puig de la Bellacasa; María José Buitrago Journal: J Clin Microbiol Date: 2014-01-29 Impact factor: 5.948
Authors: Leticia Bernal-Martínez; Laura Herrera; Clara Valero; Paula de la Cruz; Larisa Ghimpu; Ana C Mesa-Arango; Gabriela Santoni; Lidia Goterris; Rosario Millán; María José Buitrago Journal: J Fungi (Basel) Date: 2021-04-27
Authors: Luisa F López; César O Muñoz; Diego H Cáceres; Ángela M Tobón; Vladimir Loparev; Oliver Clay; Tom Chiller; Anastasia Litvintseva; Lalitha Gade; Ángel González; Beatriz L Gómez Journal: PLoS One Date: 2017-12-29 Impact factor: 3.240
Authors: A C Lehur; M Zielinski; J Pluvy; V Grégoire; S Diamantis; A Bleibtreu; C Rioux; A Picard; D Vallois Journal: BMC Infect Dis Date: 2017-05-05 Impact factor: 3.090