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A multicentre external quality assessment: A first step to standardise PCR protocols for the diagnosis of histoplasmosis and coccidioidomycosis

2023-05-11Journal article Restricted access
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dc.contributor.authorWilmes, Dunja
dc.contributor.authorHagen, Ferry
dc.contributor.authorVerissimo, Cristina
dc.contributor.authorAlanio, Alexandre
dc.contributor.authorRickerts, Volker
dc.contributor.authorBuitrago, Maria José
dc.date.accessioned2024-02-14T09:56:13Z
dc.date.available2024-02-14T09:56:13Z
dc.date.issued2023-05-11
dc.description.abstractBackground: In-house real-time PCR (qPCR) is increasingly used to diagnose the so called endemic mycoses as commercial assays are not widely available. Objectives: To compare the performance of different molecular diagnostic assays for detecting Histoplasma capsulatum and Coccidioides spp. in five European reference laboratories. Methods: Two blinded external quality assessment (EQA) panels were sent to each laboratory that performed the analysis with their in-house assays. Both panels in cluded a range of concentrations of H. capsulatum (n= 7) and Coccidioides spp. (n= 6), negative control and DNA from other fungi. Four laboratories used specific qPCRs, and one laboratory a broad-range fungal conventional PCR (cPCR) and a specific cPCR for H. capsulatum with subsequent sequencing. Results: qPCR assays were the most sensitive for the detection of H. capsulatum DNA. The lowest amount of H. capsulatum DNA detected was 1–4 fg, 0.1 pg and 10 pg for qPCRs, specific cPCR and broad-range cPCR, respectively. False positive results oc curred with high concentrations of Blastomyces dermatitidis DNA in two laboratories and with Emergomyces spp. in one laboratory. For the Coccidioides panel, the lowest amount of DNA detected was 1–16 fg by qPCRs and 10 pg with the broad-range cPCR. One labo ratory reported a false positive result by qPCR with high load of Uncinocarpus DNA Conclusion: All five laboratories were able to correctly detect H. capsulatum and Coccidioides spp. DNA and qPCRs had a better performance than specific cPCR and broad-range cPCR. EQAs may help standardise in-house molecular tests for the so called endemic mycoses improving patient management.pt_PT
dc.description.sponsorshipThis work was supported by research project PI21CIII/00007 from Spanish Fondo de Investigaciones Sanitarias of the Instituto de Salud Carlos III.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMycoses. 2023 Sep;66(9):774-786. doi: 10.1111/myc.13603. Epub 2023 May 11.pt_PT
dc.identifier.doi10.1111/myc.13603pt_PT
dc.identifier.issn0933-7407
dc.identifier.urihttp://hdl.handle.net/10400.18/9098
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherWileypt_PT
dc.relation.publisherversionhttps://onlinelibrary.wiley.com/doi/10.1111/myc.13603pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/pt_PT
dc.subjectCoccidioidespt_PT
dc.subjectKinetic Polymerase Chain Reactionpt_PT
dc.subjectHistoplasmapt_PT
dc.subjectMulticentre Trialpt_PT
dc.subjectPCRpt_PT
dc.subjectQuality Controlpt_PT
dc.subjectDiagnosispt_PT
dc.subjectInfecções Sistémicas e Zoonosespt_PT
dc.titleA multicentre external quality assessment: A first step to standardise PCR protocols for the diagnosis of histoplasmosis and coccidioidomycosispt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage786pt_PT
oaire.citation.issue9pt_PT
oaire.citation.startPage774pt_PT
oaire.citation.titleMycosespt_PT
oaire.citation.volume66pt_PT
rcaap.embargofctAcesso de acordo com política editorial da revista.pt_PT
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT

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