Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of Histoplasmosis
Lack of sensitive diagnostic tests impairs the understanding of the epidemiology of histoplasmosis, a disease whose burden is estimated to be largely underrated. Broad-range PCRs have been applied to identify fungal agents from pathology blocks, but sensitivity is variable. In this study, we compare...
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doaj-a986b4059c1f47168893d82ebda9e9842020-11-28T00:04:36ZengMDPI AGJournal of Fungi2309-608X2020-11-01631931910.3390/jof6040319Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of HistoplasmosisDunja Wilmes0Ilka McCormick-Smith1Charlotte Lempp2Ursula Mayer3Arik Bernard Schulze4Dirk Theegarten5Sylvia Hartmann6Volker Rickerts7Reference Laboratory for Cryptococcosis and Uncommon Invasive Fungal Infections, Division for Mycotic and Parasitic Agents and Mycobacteria, Robert Koch Institute, 13353 Berlin, GermanyReference Laboratory for Cryptococcosis and Uncommon Invasive Fungal Infections, Division for Mycotic and Parasitic Agents and Mycobacteria, Robert Koch Institute, 13353 Berlin, GermanyVet Med Labor GmbH, Division of IDEXX Laboratories, 71636 Ludwigsburg, GermanyVet Med Labor GmbH, Division of IDEXX Laboratories, 71636 Ludwigsburg, GermanyDepartment of Medicine A, Hematology, Oncology and Pulmonary Medicine, University Hospital Muenster, 48149 Muenster, GermanyInstitute of Pathology, University Hospital Essen, University Duisburg-Essen, 45147 Essen, GermanySenckenberg Institute for Pathology, Johann Wolfgang Goethe University Frankfurt, 60323 Frankfurt am Main, GermanyReference Laboratory for Cryptococcosis and Uncommon Invasive Fungal Infections, Division for Mycotic and Parasitic Agents and Mycobacteria, Robert Koch Institute, 13353 Berlin, GermanyLack of sensitive diagnostic tests impairs the understanding of the epidemiology of histoplasmosis, a disease whose burden is estimated to be largely underrated. Broad-range PCRs have been applied to identify fungal agents from pathology blocks, but sensitivity is variable. In this study, we compared the results of a specific <i>Histoplasma</i> qPCR (<i>H</i>. qPCR) with the results of a broad-range qPCR (28S qPCR) on formalin-fixed, paraffin-embedded (FFPE) tissue specimens from patients with proven fungal infections (<i>n</i> = 67), histologically suggestive of histoplasmosis (<i>n</i> = 36) and other mycoses (<i>n</i> = 31). The clinical sensitivity for histoplasmosis of the <i>H</i>. qPCR and the 28S qPCR was 94% and 48.5%, respectively. Samples suggestive for other fungal infections were negative with the <i>H</i>. qPCR. The 28S qPCR did not amplify DNA of <i>Histoplasma</i> in FFPE in these samples, but could amplify DNA of <i>Emergomyces</i> (<i>n</i> = 1) and <i>Paracoccidioides</i> (<i>n</i> = 2) in three samples suggestive for histoplasmosis but negative in the <i>H.</i> qPCR. In conclusion, amplification of <i>Histoplasma</i> DNA from FFPE samples is more sensitive with the <i>H.</i> qPCR than with the 28S qPCR. However, the 28S qPCR identified DNA of other fungi in <i>H</i>. qPCR-negative samples presenting like histoplasmosis, suggesting that the combination of both assays may improve the diagnosis.https://www.mdpi.com/2309-608X/6/4/319<i>Histoplasma</i> qPCRbroad-range qPCRformalin-fixed paraffin-embedded (FFPE) sampleshistoplasmosis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Dunja Wilmes Ilka McCormick-Smith Charlotte Lempp Ursula Mayer Arik Bernard Schulze Dirk Theegarten Sylvia Hartmann Volker Rickerts |
spellingShingle |
Dunja Wilmes Ilka McCormick-Smith Charlotte Lempp Ursula Mayer Arik Bernard Schulze Dirk Theegarten Sylvia Hartmann Volker Rickerts Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of Histoplasmosis Journal of Fungi <i>Histoplasma</i> qPCR broad-range qPCR formalin-fixed paraffin-embedded (FFPE) samples histoplasmosis |
author_facet |
Dunja Wilmes Ilka McCormick-Smith Charlotte Lempp Ursula Mayer Arik Bernard Schulze Dirk Theegarten Sylvia Hartmann Volker Rickerts |
author_sort |
Dunja Wilmes |
title |
Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of Histoplasmosis |
title_short |
Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of Histoplasmosis |
title_full |
Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of Histoplasmosis |
title_fullStr |
Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of Histoplasmosis |
title_full_unstemmed |
Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of Histoplasmosis |
title_sort |
detection of histoplasma dna from tissue blocks by a specific and a broad-range real-time pcr: tools to elucidate the epidemiology of histoplasmosis |
publisher |
MDPI AG |
series |
Journal of Fungi |
issn |
2309-608X |
publishDate |
2020-11-01 |
description |
Lack of sensitive diagnostic tests impairs the understanding of the epidemiology of histoplasmosis, a disease whose burden is estimated to be largely underrated. Broad-range PCRs have been applied to identify fungal agents from pathology blocks, but sensitivity is variable. In this study, we compared the results of a specific <i>Histoplasma</i> qPCR (<i>H</i>. qPCR) with the results of a broad-range qPCR (28S qPCR) on formalin-fixed, paraffin-embedded (FFPE) tissue specimens from patients with proven fungal infections (<i>n</i> = 67), histologically suggestive of histoplasmosis (<i>n</i> = 36) and other mycoses (<i>n</i> = 31). The clinical sensitivity for histoplasmosis of the <i>H</i>. qPCR and the 28S qPCR was 94% and 48.5%, respectively. Samples suggestive for other fungal infections were negative with the <i>H</i>. qPCR. The 28S qPCR did not amplify DNA of <i>Histoplasma</i> in FFPE in these samples, but could amplify DNA of <i>Emergomyces</i> (<i>n</i> = 1) and <i>Paracoccidioides</i> (<i>n</i> = 2) in three samples suggestive for histoplasmosis but negative in the <i>H.</i> qPCR. In conclusion, amplification of <i>Histoplasma</i> DNA from FFPE samples is more sensitive with the <i>H.</i> qPCR than with the 28S qPCR. However, the 28S qPCR identified DNA of other fungi in <i>H</i>. qPCR-negative samples presenting like histoplasmosis, suggesting that the combination of both assays may improve the diagnosis. |
topic |
<i>Histoplasma</i> qPCR broad-range qPCR formalin-fixed paraffin-embedded (FFPE) samples histoplasmosis |
url |
https://www.mdpi.com/2309-608X/6/4/319 |
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