{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T01:50:13Z","timestamp":1780451413943,"version":"3.54.1"},"reference-count":31,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2024,6,25]],"date-time":"2024-06-25T00:00:00Z","timestamp":1719273600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100014555","name":"Murdoch Children\u2019s Research Institute","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100014555","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Pleural empyema is a serious complication of pneumonia in children. Negative bacterial cultures commonly impede optimal antibiotic therapy. To improve bacterial identification, we developed a molecular assay and evaluated its performance compared with bacterial culture. Our multiplex-quantitative PCR to detect <jats:italic>Streptococcus pneumoniae<\/jats:italic>, <jats:italic>Streptococcus pyogenes<\/jats:italic>, <jats:italic>Staphylococcus aureus<\/jats:italic> and <jats:italic>Haemophilus influenzae<\/jats:italic> was assessed using bacterial genomic DNA and laboratory-prepared samples (n = 267). To evaluate clinical performance, we conducted the Molecular Assessment of Thoracic Empyema (MATE) observational study, enrolling children hospitalised with empyema. Pleural fluids were tested by bacterial culture and multiplex-qPCR, and performance determined using a study gold standard. We determined clinical sensitivity and time-to-organism-identification to assess the potential of the multiplex-qPCR to reduce the duration of empiric untargeted antibiotic therapy. Using spiked samples, the multiplex-qPCR demonstrated 213\/215 (99.1%) sensitivity and 52\/52 (100%) specificity for all organisms. During May 2019\u2013March 2023, 100 children were enrolled in the MATE study; median age was 3.9 years (IQR 2\u20135.6). A bacterial pathogen was identified in 90\/100 (90%) specimens by multiplex-qPCR, and 24\/100 (24%) by bacterial culture (<jats:italic>P<\/jats:italic> &lt;0.001). Multiplex-qPCR identified a bacterial cause in 68\/76 (90%) culture-negative specimens. <jats:italic>S<\/jats:italic>. <jats:italic>pneumoniae<\/jats:italic> was the most common pathogen, identified in 67\/100 (67%) specimens. We estimate our multiplex-qPCR would have reduced the duration of untargeted antibiotic therapy in 61% of cases by a median 20 days (IQR 17.5\u201323, range 1\u201355). Multiplex-qPCR significantly increased pathogen detection compared with culture and may allow for reducing the duration of untargeted antibiotic therapy.<\/jats:p>","DOI":"10.1371\/journal.pone.0304861","type":"journal-article","created":{"date-parts":[[2024,6,25]],"date-time":"2024-06-25T19:15:16Z","timestamp":1719342916000},"page":"e0304861","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Evaluation of a multiplex-qPCR for paediatric pleural empyema\u2014An observational study in hospitalised 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