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2023-05-11Zeitschriftenartikel
A multicentre external quality assessment: A first step to standardise PCR protocols for the diagnosis of histoplasmosis and coccidioidomycosis
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.accessioned2023-11-15T15:06:48Z
dc.date.available2023-11-15T15:06:48Z
dc.date.issued2023-05-11none
dc.identifier.other10.1111/myc.13603
dc.identifier.urihttp://edoc.rki.de/176904/11351
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 included 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 occurred 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 laboratory 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.eng
dc.language.isoengnone
dc.publisherRobert Koch-Institut
dc.rights(CC BY-NC-ND 3.0 DE) Namensnennung - Nicht-kommerziell - Keine Bearbeitung 3.0 Deutschlandger
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/de/
dc.subjectCoccidiodeseng
dc.subjectHistoplasmaeng
dc.subjectkinetic polymerase chain reactioneng
dc.subjectmulticentre trialeng
dc.subjectPCReng
dc.subjectquality controleng
dc.subject.ddc610 Medizin und Gesundheitnone
dc.titleA multicentre external quality assessment: A first step to standardise PCR protocols for the diagnosis of histoplasmosis and coccidioidomycosisnone
dc.typearticle
dc.identifier.urnurn:nbn:de:0257-176904/11351-1
dc.type.versionpublishedVersionnone
local.edoc.container-titleMycosesnone
local.edoc.container-issn1439-0507none
local.edoc.pages13none
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttps://onlinelibrary.wiley.com/journal/14390507none
local.edoc.container-publisher-nameWileynone
local.edoc.container-volume66none
local.edoc.container-issue12none
local.edoc.container-reportyear2023none
local.edoc.container-firstpage774none
local.edoc.container-lastpage786none
dc.description.versionPeer Reviewednone

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