Evaluation of a multiplex-qPCR for paediatric pleural empyema-An observational study in hospitalised children

• Published in: PloS one 2024-06, Vol.19 (6), p.e0304861
• Main Authors: Jacobson, Jonathan, Fabri, Loraine, Osowicki, Joshua, Shanthikumar, Shivanthan, Costa, Anna-Maria, Ortika, Belinda, Wee-Hee, Ashleigh, Pragassen, Michelle, Gatt, Cassandra, Gonis, Gena, Nguyen, Cattram, Rozen, Thomas, Teague, Warwick, Buttery, Jim, Clifford, Vanessa, Mulholland, Kim, Steer, Andrew, Ranganathan, Sarath, Daley, Andrew, Dunne, Eileen, Satzke, Catherine
• Format: Article
• Language: English
• Subjects: Anti-Bacterial Agents - therapeutic use; Antibiotics; Bacteria; Bacterial pneumonia; Child; Child, Preschool; Children; Consent; DNA, Bacterial - genetics; Empyema; Empyema, Pleural - diagnosis; Empyema, Pleural - drug therapy; Empyema, Pleural - microbiology; Ethylenediaminetetraacetic acid; Female; Haemophilus influenzae; Haemophilus influenzae - genetics; Haemophilus influenzae - isolation & purification; Hospitalization; Humans; Infant; Male; Medical records; Multiplex Polymerase Chain Reaction - methods; Multiplexing; Observational studies; Pathogens; Patients; Pediatrics; Performance evaluation; Pneumonia; Sensitivity analysis; Sensitivity and Specificity; Staphylococcus aureus; Staphylococcus aureus - drug effects; Staphylococcus aureus - genetics; Staphylococcus aureus - isolation & purification; Streptococcus infections; Streptococcus pneumoniae; Streptococcus pneumoniae - genetics; Streptococcus pneumoniae - isolation & purification; Streptococcus pyogenes; Streptococcus pyogenes - genetics; Streptococcus pyogenes - isolation & purification; Therapy; Thorax
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0304861

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 Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus and Haemophilus influenzae 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-March 2023, 100 children were enrolled in the MATE study; median age was 3.9 years (IQR 2-5.6). A bacterial pathogen was identified in 90/100 (90%) specimens by multiplex-qPCR, and 24/100 (24%) by bacterial culture (P