{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T05:52:24Z","timestamp":1785909144271,"version":"3.56.0"},"reference-count":29,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pediatr."],"abstract":"<jats:sec>\n                    <jats:title>Purpose<\/jats:title>\n                    <jats:p>Infectious diseases constitute the predominant cause of fever of unknown origin (FUO). Conventional microbiological testing is limited by prolonged turnaround times, susceptibility to host\/environmental interference, low detection sensitivity, and limited capacity to identify rare pathogens. Metagenomic next-generation sequencing (mNGS) enables parallel broad-spectrum screening for microbial agents. This study aimed to investigate the clinical utility of mNGS in children presenting with FUO, to generate descriptive observational data on pathogen detection and temporally associated anti-infective regimen adjustments.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>This retrospective single-center analysis enrolled 41 hospitalized children diagnosed with FUO who underwent mNGS testing at the Department of Infectious Diseases, Affiliated Children's Hospital of Shandong University, from June 2022 to July 2025. Initially, all patients underwent comprehensive routine systemic evaluations. For cases where fever persisted despite conventional testing and an infectious etiology was highly suspected, or where there was a poor therapeutic response to empirical anti-infective treatments, mNGS was subsequently performed. All specimens submitted for testing were sterile body fluids. Each sample was divided into two aliquots: one was subjected to conventional microbiological testing (including culture, smear microscopy, and PCR), while the other was cryopreserved for mNGS analysis. The performance of pathogen detection was compared between mNGS and conventional testing modalities using paired specimen data.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      In this study, we analyzed 41 pediatric cases, which included three types of specimens: blood, cerebrospinal fluid (CSF), and tissue fluid (comprising deep pus, postoperative drainage fluid, subdural effusion, and aspirated fluid from the mass). mNGS identified 30 microbial isolates from 20 patients, which included bacteria, viruses, fungi, and mycoplasmas; of these, 17 isolates were ultimately confirmed as causative pathogens. No statistically significant differences in positivity rates were observed between mNGS and conventional assays, as indicated by paired 2\u2009\u00d7\u20092 contingency tables (all\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&amp;gt;\u20090.05).The present study also recorded changes to antimicrobial regimens that occurred after pathogen identification by mNGS testing, including adjuvant antiviral therapy for 4 patients, antimicrobial escalation for 6 patients, antimicrobial de-escalation for 2 patients, and comprehensive regimen modifications for an additional 5 patients.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>We analyzed a targeted pediatric FUO subgroup, and the overall pathogen detection positivity rate showed no statistical difference between mNGS and routine microbial testing. Accordingly, mNGS cannot currently replace standard workflows or routinely screen all FUO children. The two testing methods exhibited complementary pathogen detection spectra. mNGS may act as an auxiliary tool for complicated infectious cases with negative conventional test results. This study generates descriptive observational data on pathogen identification and temporally associated anti-infective regimen adjustments in a selected cohort of FUO children. Further prospective studies with larger sample sizes are required to validate these findings.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fped.2026.1868060","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T05:39:45Z","timestamp":1785908385000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Clinical application value of metagenomic next-generation sequencing in children with fever of unknown origin"],"prefix":"10.3389","volume":"14","author":[{"given":"Shihua","family":"Liu","sequence":"first","affiliation":[{"name":"Department of Infectious Diseases, Children's Hospital Affiliated to Shandong University (Jinan Children's Hospital)","place":["Jinan, Shandong Province, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaoning","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Children's Hospital Affiliated to Shandong University (Jinan Children's Hospital)","place":["Jinan, Shandong Province, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Children's Hospital Affiliated to Shandong University (Jinan Children's Hospital)","place":["Jinan, Shandong Province, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"ofaa132","DOI":"10.1093\/ofid\/ofaa132","article-title":"Fever and fever of unknown origin: review, recent advances, and lingering dogma","volume":"7","author":"Wright","year":"2020","journal-title":"Open Forum Infect Dis"},{"key":"B2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1097\/00005792-196102000-00001","article-title":"Fever of unexplained origin: report on 100 cases","volume":"40","author":"Petersdorf","year":"1961","journal-title":"Medicine (Baltimore)"},{"key":"B3","doi-asserted-by":"publisher","first-page":"257","DOI":"10.3760\/cma.j.cn311365-20260120-00026","article-title":"Expert consensus on the diagnosis and management of fever of unknown origin (2026 edition)","volume":"44","author":"Zhang","year":"2026","journal-title":"Chin J Infect Dis"},{"key":"B4","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1056\/NEJMra2111003","article-title":"Fever of unknown origin","volume":"386","author":"Haidar","year":"2022","journal-title":"N Engl J Med"},{"key":"B5","doi-asserted-by":"publisher","first-page":"e080847","DOI":"10.1136\/bmj-2024-080847","article-title":"Fever of unknown origin","volume":"388","author":"Wright","year":"2025","journal-title":"Br Med J"},{"key":"B6","doi-asserted-by":"publisher","first-page":"20","DOI":"10.3390\/children11010020","article-title":"Update on fever of unknown origin in children: focus on etiologies and clinical approach","volume":"11","author":"Trapani","year":"2024","journal-title":"Children (Basel)"},{"key":"B7","doi-asserted-by":"publisher","first-page":"102824","DOI":"10.1016\/j.jiph.2025.102824","article-title":"Epidemiology and diagnostic challenges of fever of unknown origin (FUO) among adults: a multicenter retrospective study in northern Italy","volume":"18","author":"De Pascali","year":"2025","journal-title":"J Infect Public Health"},{"key":"B8","doi-asserted-by":"publisher","first-page":"452","DOI":"10.1186\/s12879-023-08436-0","article-title":"Structured diagnostic scheme clinical experience sharing: a prospective study of 320 cases of fever of unknown origin in a tertiary hospital in north China","volume":"23","author":"Jiang","year":"2023","journal-title":"BMC Infect Dis"},{"key":"B9","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ijid.2019.10.020","article-title":"Metagenomic next-generation sequencing helped diagnose scrub typhus without eschar: a case report","volume":"90","author":"Wu","year":"2020","journal-title":"Int J Infect Dis"},{"key":"B10","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1080\/22221751.2020.1725399","article-title":"RNA Based mNGS approach identifies a novel human coronavirus from two individual pneumonia cases in 2019 Wuhan outbreak","volume":"9","author":"Chen","year":"2020","journal-title":"Emerging Microbes Infect"},{"key":"B11","doi-asserted-by":"publisher","first-page":"323","DOI":"10.1016\/j.jinf.2019.01.003","article-title":"Evaluation of next-generation sequencing for the pathogenic diagnosis of children brain abscesses","volume":"78","author":"Hu","year":"2019","journal-title":"J Infect"},{"key":"B12","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1007\/s10096-025-05289-0","article-title":"Clinical application of metagenomic next-generation sequencing (mNGS) in children with suspected bloodstream infection","volume":"45","author":"Pan","year":"2026","journal-title":"Eur J Clin Microbiol Infect Dis"},{"key":"B13","doi-asserted-by":"publisher","first-page":"1743423","DOI":"10.3389\/fpubh.2025.1743423","article-title":"Clinical and epidemiological characteristics of cat scratch disease in children from southwestern China: a retrospective analysis of mNGS-confirmed cases","volume":"13","author":"Lai","year":"2026","journal-title":"Front Public Health"},{"key":"B14","doi-asserted-by":"publisher","first-page":"601","DOI":"10.3390\/pathogens14060601","article-title":"Deploying metagenomics to characterize microbial pathogens during outbreak of acute febrile illness among children in Tanzania","volume":"14","author":"Mziray","year":"2025","journal-title":"Pathogens (Basel, Switzerland)"},{"key":"B15","doi-asserted-by":"publisher","first-page":"1189","DOI":"10.3855\/jidc.20668","article-title":"Detection of pathogens and epidemiological characteristics of community-acquired pneumonia in children using metagenomic next-generation sequencing","volume":"19","author":"Deng","year":"2025","journal-title":"J Infect Dev Ctries"},{"key":"B16","doi-asserted-by":"publisher","first-page":"1165432","DOI":"10.3389\/fcimb.2023.1165432","article-title":"High specificity of metagenomic next-generation sequencing using protected bronchial brushing sample in diagnosing pneumonia in children","volume":"13","author":"Zhang","year":"2023","journal-title":"Front Cell Infect Microbiol"},{"key":"B17","doi-asserted-by":"publisher","first-page":"1368165","DOI":"10.3389\/fcimb.2024.1368165","article-title":"Clinical features of pediatric mucormycosis: role of metagenomic next generation sequencing in diagnosis","volume":"14","author":"Zhang","year":"2024","journal-title":"Front Cell Infect Microbiol"},{"key":"B18","doi-asserted-by":"publisher","first-page":"231","DOI":"10.1186\/s13052-024-01804-9","article-title":"Duration of onset, body temperature and C-reactive protein can be used to predict the results of pus culture in children with acute osteomyelitis of long bones","volume":"50","author":"Jia","year":"2024","journal-title":"Ital J Pediatr"},{"key":"B19","doi-asserted-by":"publisher","first-page":"326","DOI":"10.1093\/jpids\/piz032","article-title":"Prospective cohort study of next-generation sequencing as a diagnostic modality for unexplained encephalitis in children","volume":"9","author":"Haston","year":"2020","journal-title":"J Pediatric Infect Dis Soc"},{"key":"B20","doi-asserted-by":"publisher","first-page":"e0218318","DOI":"10.1371\/journal.pone.0218318","article-title":"Metagenomic next-generation sequencing of samples from pediatric febrile illness in Tororo, Uganda","volume":"14","author":"Ramesh","year":"2019","journal-title":"PLoS One"},{"key":"B21","doi-asserted-by":"publisher","first-page":"e049840","DOI":"10.1136\/bmjopen-2021-049840","article-title":"Fever of unknown origin (FUO) in children: a single-centre experience from Beijing, China","volume":"12","author":"Hu","year":"2022","journal-title":"BMJ open"},{"key":"B22","doi-asserted-by":"publisher","first-page":"e00053","DOI":"10.1128\/CMR.00053-19","article-title":"Performance and application of 16S rRNA gene cycle sequencing for routine identification of Bacteria in the clinical microbiology laboratory","volume":"33","author":"Church","year":"2020","journal-title":"Clin Microbiol Rev"},{"key":"B23","doi-asserted-by":"publisher","first-page":"36","DOI":"10.56964\/pidspj20171801006","article-title":"Fever of unknown origin in children: a five-year review","volume":"18","author":"Cabanalan-Rivera","year":"2017","journal-title":"Pediatric Infectious Disease Society of the Philippines Journal"},{"key":"B24","doi-asserted-by":"publisher","first-page":"196","DOI":"10.1016\/j.jmoldx.2019.10.007","article-title":"Retrospective validation of a metagenomic sequencing protocol for combined detection of RNA and DNA viruses using respiratory samples from pediatric patients","volume":"22","author":"van Boheemen","year":"2020","journal-title":"J Mol Diagn"},{"key":"B25","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1016\/j.jviromet.2015.05.010","article-title":"Evaluation of convenient pretreatment protocols for RNA virus metagenomics in serum and tissue samples","volume":"222","author":"Rosseel","year":"2015","journal-title":"J Virol Methods"},{"key":"B26","doi-asserted-by":"publisher","first-page":"2395","DOI":"10.2147\/IDR.S403851","article-title":"The impact of mNGS technology in the etiological diagnosis of severe pneumonia in children during the epidemic of COVID-19","volume":"16","author":"Yu","year":"2023","journal-title":"Infect Drug Resist"},{"key":"B27","doi-asserted-by":"publisher","first-page":"e00794","DOI":"10.1128\/JCM.00794-20","article-title":"Plasma metagenomic next-generation sequencing assay for identifying pathogens: a retrospective review of test utilization in a large children's hospital","volume":"58","author":"Niles","year":"2020","journal-title":"J Clin Microbiol"},{"key":"B28","doi-asserted-by":"publisher","first-page":"371","DOI":"10.1080\/14737159.2021.1906652","article-title":"Rapid metagenomics for diagnosis of bloodstream and respiratory tract nosocomial infections: current status and future prospects","volume":"21","author":"Moragues-Solanas","year":"2021","journal-title":"Expert Rev Mol Diagn"},{"key":"B29","doi-asserted-by":"publisher","first-page":"778","DOI":"10.1093\/cid\/cix881","article-title":"Understanding the promises and hurdles of metagenomic next-generation sequencing as a diagnostic tool for infectious diseases","volume":"66","author":"Simner","year":"2018","journal-title":"Clin Infect Dis"}],"container-title":["Frontiers in Pediatrics"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fped.2026.1868060\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T05:39:46Z","timestamp":1785908386000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fped.2026.1868060\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,5]]},"references-count":29,"alternative-id":["10.3389\/fped.2026.1868060"],"URL":"https:\/\/doi.org\/10.3389\/fped.2026.1868060","relation":{},"ISSN":["2296-2360"],"issn-type":[{"value":"2296-2360","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,5]]},"article-number":"1868060"}}