{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,17]],"date-time":"2026-10-17T19:58:24Z","timestamp":1792267104768,"version":"4.4.0"},"reference-count":27,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2025,2,19]],"date-time":"2025-02-19T00:00:00Z","timestamp":1739923200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,2,19]],"date-time":"2025-02-19T00:00:00Z","timestamp":1739923200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2025,2,19]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      The aim of this study was to identify parameters influencing DNA extraction and PCR amplification efficiencies in an attempt to standardize Mucorales qPCR. The Fungal PCR Initiative Mucorales Laboratory Working Group distributed two panels of simulated samples to 26 laboratories: Panel A (six sera spiked with Mucorales DNA and one negative control serum) and Panel B (six Mucorales DNA extracts). Panel A underwent DNA extraction in each laboratory according to the local procedure and were sent to a central laboratory for testing using three different qPCR techniques: one in-house qPCR assay and two commercial assays (MucorGenius and Fungiplex). Panel B DNA extracts were PCR amplified in each laboratory using local procedures: nine in-house qPCR assays and two commercial kits (MucorGenius and MycoGENIE). All data were compiled and anonymously analyzed at the central laboratory. For Panel A, a total of six different automated platforms and five manual extraction methods were used. Positive rates were 64%, 70%, and 89%, for the MucorGenius, Fungiplex, and the in-house qPCR assay, respectively. Using a large volume of serum for DNA extraction provided the highest analytical sensitivity (82.5% for 1 mL compared with 62.7% for smaller volumes,\n                      <jats:italic>P<\/jats:italic>\n                      &lt; 0.01). For Panel B, five in-house qPCR assays and two commercial kits had &gt;78% positivity. Using larger PCR input volumes (\u22657 \u00b5L) was associated with the highest sensitivity at 95.5% compared to 58.3% when lower input volumes were used (\n                      <jats:italic>P<\/jats:italic>\n                      &lt; 0.01). Using larger sample volumes for nucleic acid extraction and DNA template volumes for PCR amplification significantly improves the performance of Mucorales qPCR when testing serum.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>Mucormycosis is a life-threatening mold infection affecting immunosuppressed patients but also other patients with diabetes or trauma. Better survival is linked to shorter delays in diagnosis and treatment initiation. Detection of Mucorales-free DNA in serum or plasma using quantitative PCR allows a prompt diagnosis and earlier treatment. Several techniques and protocols of quantitative Mucorales PCR are used in Europe, and improving performance remains a common objective of laboratories participating in the fungal PCR Initiative Working Group. This study, which combined results from 26 laboratories in Europe, showed that the main parameters underpinning sensitivity are the preanalytical variables (volume of serum used for DNA extraction and DNA template volume), irrespective of the extraction platforms and qPCR assay\/platform.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/jcm.01525-24","type":"journal-article","created":{"date-parts":[[2024,12,31]],"date-time":"2024-12-31T08:00:38Z","timestamp":1735632038000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":15,"title":["Interlaboratory assays from the fungal PCR Initiative and the Modimucor Study Group to improve qPCR detection of Mucorales DNA in serum: one more step toward standardization"],"prefix":"10.1128","volume":"63","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9262-1127","authenticated-orcid":false,"given":"Steffi","family":"Rocchi","sequence":"first","affiliation":[{"name":"Chrono-environnement UMR6249, CNRS, University of Franche-Comt\u00e9","place":["Besan\u00e7on, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emeline","family":"Scherer","sequence":"additional","affiliation":[{"name":"Chrono-environnement UMR6249, CNRS, University of Franche-Comt\u00e9","place":["Besan\u00e7on, France"]},{"name":"Parasitology\u2013Mycology Department, Besan\u00e7on University Hospital","place":["Besan\u00e7on, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3056-4205","authenticated-orcid":true,"given":"P. Lewis","family":"White","sequence":"additional","affiliation":[{"name":"Public Health Wales Mycology Reference Laboratory, University Hospital of Wales","place":["Cardiff, United Kingdom"]},{"name":"Division of Infection and Immunity, Centre for Trials Research","place":["Cardiff, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Audrey","family":"Guitton","sequence":"additional","affiliation":[{"name":"Chrono-environnement UMR6249, CNRS, University of Franche-Comt\u00e9","place":["Besan\u00e7on, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre","family":"Alanio","sequence":"additional","affiliation":[{"name":"Laboratoire de parasitologie-mycologie, AP-HP, H\u00f4pital Saint-Louis","place":["Paris, France"]},{"name":"Mycology Department, Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, National Reference Center for Invasive Mycoses and Antifungals, Translational Mycology research group","place":["Paris, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fran\u00e7oise","family":"Botterel","sequence":"additional","affiliation":[{"name":"Unit\u00e9 de Parasitologie-Mycologie, D\u00e9partement de Pr\u00e9vention, Diagnostic et Traitement des Infections, CHU Henri Mondor, Assistance Publique des H\u00f4pitaux de Paris (APHP)","place":["Cr\u00e9teil, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4721-2740","authenticated-orcid":true,"given":"Marie Elisabeth","family":"Bougnoux","sequence":"additional","affiliation":[{"name":"Parasitology-Mycology Unit, Necker Enfants Malades Hospital, APHP","place":["Paris, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Jos\u00e9","family":"Buitrago","sequence":"additional","affiliation":[{"name":"Mycology Reference Laboratory, Centro Nacional de Microbiologia, Instituto de Salud Carlos III","place":["Madrid, Spain"]},{"name":"CIBERINFEC, ISCIII-CIBER de Enfermedades Infecciosas, Instituto de Salud Carlos III","place":["Madrid, Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Massimo","family":"Cogliati","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences for Health, Universit\u00e0 degli Studi di Milano","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marjorie","family":"Cornu","sequence":"additional","affiliation":[{"name":"Universit\u00e9 de Lille, Inserm U1285, CHU Lille, Laboratoire de Parasitologie-Mycologie, CNRS, UMR 8576, UGSF\u2014Unite\u0301 de Glycobiologie Structurale et Fonctionnelle","place":["Lille, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Celine","family":"Damiani","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie et Mycologie M\u00e9dicales, Centre de Biologie Humaine, CHU Amiens Picardie, Agents Infectieux, R\u00e9sistance et Chimioth\u00e9rapie 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