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  • A multicentre external quality assessment: A first step to standardise PCR protocols for the diagnosis of histoplasmosis and coccidioidomycosis
    Publication . Wilmes, Dunja; Hagen, Ferry; Verissimo, Cristina; Alanio, Alexandre; Rickerts, Volker; Buitrago, Maria José
    Background: 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.
    2023-05-11Journal article Restricted access Show more
  • Rapid cross-border emergence of NDM-5-producing Escherichia coli in the European Union/European Economic Area, 2012 to June 2022
    Publication . Linkevicius, Marius; Bonnin, Rémy A.; Alm, Erik; Svartström, Olov; Apfalter, Petra; Hartl, Rainer; Hasman, Henrik; Roer, Louise; Räisänen, Kati; Dortet, Laurent; Pfennigwerth, Niels; Hans, Jörg B.; Tóth, Ákos; Buzgó, Lilla; Cormican, Martin; Delappe, Niall; Monaco, Monica; Giufrè, Maria; Hendrickx, Antoni P.A.; Samuelsen, Ørjan; Pöntinen, Anna K.; Caniça, Manuela; Manageiro, Vera; Oteo-Iglesias, Jesús; Pérez-Vázquez, María; Westmo, Karin; Mäkitalo, Barbro; Palm, Daniel; Monnet, Dominique L.; Kohlenberg, Anke
    Whole genome sequencing data of 874 Escherichia coli isolates carrying bla NDM-5 from 13 European Union/European Economic Area countries between 2012 and June 2022 showed the predominance of sequence types ST167, ST405, ST410, ST361 and ST648, and an increasing frequency of detection. Nearly a third (30.6%) of these isolates were associated with infections and more than half (58.2%) were predicted to be multidrug-resistant. Further spread of E. coli carrying bla NDM-5 would leave limited treatment options for serious E. coli infections.
    2023-05-11Journal article Open access Show more