Comparison of PCR protocols for detecting Histoplasma capsulatum DNA through a multicenter study |
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| Authors: | María José Buitrago Cristina Elena Canteros Guadalupe Frías De León Ángel González Manoel Marques-Evangelista De Oliveira César O Muñoz José Antonio Ramirez Adriana Isabel Toranzo Rosely Zancope-Oliveira Manuel Cuenca-Estrella |
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| Institution: | 1. Servicio de Micología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra Majadahonda-Pozuelo, Km 2, 28220 Majadahonda (Madrid), Spain;2. Unidad de Micología Médica y Experimental, Corporación para Investigaciones Biológicas (CIB) y Escuela de Microbiología, Universidad de Antioquia, Medellín, Colombia;3. Setor de Imunodiagnóstico do Laboratório de Micologia do Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil;4. Servicio Micosis Profundas, Departamento Micología, Instituto Nacional de Enfermedades Infecciosas, ANLIS “Dr. Carlos G. Malbrán”, Buenos Aires, Argentina;5. Departamento de Microbiología y Parasitología, UNAM, México D.F., Mexico;6. División de Investigación, Hospital Juárez de México, Av. Politécnico Nacional 5160, Colonia Magdalena de las Salinas, Delegación Gustavo A. Madero, C.P. 07760, México D.F., Mexico |
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| Abstract: | BackgroundA 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.AimsThe objective was to assess the accuracy of different PCR protocols to detect H. capsulatum DNA.MethodsSeven different PCR protocols were tested. They were based on PCR techniques and used unicopy and multicopy targets.ResultsMost of these protocols (4/7) were able to detect the smallest amounts of fungal DNA (102 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.ConclusionsAll 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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| Keywords: | Histoplasmosis Multicenter PCR Diagnosis Histoplasmosis Multicéntrico PCR Diagnóstico |
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