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Med."],"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:p>In immunocompromised pediatric patients, diagnosing invasive pulmonary aspergillosis (IPA) poses a significant challenge. Next-Generation Sequencing (NGS) shows promise for detecting fungal DNA but lacks standardization. This study aims to advance towards clinical evaluation of liquid biopsy NGS for <jats:italic>Aspergillus<\/jats:italic> detection, through an evaluation of wet-lab procedures and computational analysis. Our findings support using both CHM13v2.0 and GRCh38.p14 in host-read mapping to reduce fungal false-positives. We demonstrate the sensitivity of our custom kraken2 database, cRE.21, in detecting <jats:italic>Aspergillus<\/jats:italic> species. Additionally, cell-free DNA sequencing shows superior performance to whole-cell DNA sequencing by recovering higher fractions of fungal DNA in lung fluid (bronchoalveolar lavage [BAL] fluid) and plasma samples from pediatric patients with probable IPA. In a proof-of-principle, <jats:italic>A. fumigatus<\/jats:italic> was identified in 5 out of 7 BAL fluid samples and 3 out of 5 plasma samples. This optimized workflow can advance fungal-NGS research and represents a step towards enhancing diagnostic certainty by enabling more sensitive and accurate species-level diagnosis of IPA in immunocompromised patients.<\/jats:p>","DOI":"10.1038\/s41525-025-00482-8","type":"journal-article","created":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T11:36:53Z","timestamp":1742211413000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["NGS-based Aspergillus detection in plasma and lung lavage of children with invasive pulmonary aspergillosis"],"prefix":"10.1038","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6656-2151","authenticated-orcid":false,"given":"Emmy","family":"Wesdorp","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura","family":"Rotte","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7956-5116","authenticated-orcid":false,"given":"Li-Ting","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Myrthe","family":"Jager","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolle","family":"Besselink","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlo","family":"Vermeulen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5622-1916","authenticated-orcid":false,"given":"Ferry","family":"Hagen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tjomme","family":"van der Bruggen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caroline","family":"Lindemans","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tom","family":"Wolfs","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Louis","family":"Bont","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0828-3477","authenticated-orcid":false,"given":"Jeroen","family":"de Ridder","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,3,17]]},"reference":[{"key":"482_CR1","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1016\/j.ejca.2019.08.008","volume":"121","author":"EAH Loeffen","year":"2019","unstructured":"Loeffen, E. 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C.L. has received financial support from Pfizer for educational purposes. L.B. has regular interaction with pharmaceutical and other industrial partners. L.B. has not received personal fees or other personal benefits, but UMCU has received significant funding (>\u20ac100,000 per industrial partner) for investigator-initiated studies from AstraZeneca, Sanofi, Janssen, Pfizer, MSD, and MeMed Diagnostics, as well as major funding from the Bill and Melinda Gates Foundation and through public-private partnerships such as the IMI-funded RESCEU and PROMISE projects, involving partners GSK, Novavax, Janssen, AstraZeneca, Pfizer, and Sanofi, along with substantial funding from Julius Clinical for participation in clinical studies sponsored by AstraZeneca, Merck, and Pfizer, and minor funding (\u20ac1,000-25,000 per industrial partner) for consultation, DSMB membership, or invited lectures by Ablynx, Bavaria Nordic, GSK, Novavax, Pfizer, Moderna, AstraZeneca, MSD, Sanofi, and Janssen. L.B. is the founding chairman of the ReSViNET Foundation. J.d.R. is cofounder and CTO of Cyclomics, a genomics company. L.R., L.C., M.J., N.B., C.V., T.v.d.B. and T.W. declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"24"}}