title: 16S pan-bacterial PCR can accurately identify patients with ventilator-associated pneumonia.
abstract: Ventilator-associated pneumonia (VAP) remains a challenge to intensive care units, with secure diagnosis relying on microbiological cultures that take up to 72 hours to provide a result. We sought to derive and validate a novel, real-time 16S rRNA gene PCR for rapid exclusion of VAP. Bronchoalveolar lavage (BAL) was obtained from two independent cohorts of patients with suspected VAP. Patients were recruited in a 2-centre derivation cohort and a 12-centre confirmation cohort. Confirmed VAP was defined as growth of >104colony forming units/ml on semiquantitative culture and compared with a 16S PCR assay. Samples were tested from 67 patients in the derivation cohort, 10 (15%) of whom had confirmed VAP. Using cycles to cross threshold (Ct) values as the result of the 16S PCR test, the area under the receiver operating characteristic (ROC) curve (AUROC) was 0.94 (95% CI 0.86 to 1.0, p<0.0001). Samples from 92 patients were available from the confirmation cohort, 26 (28%) of whom had confirmed VAP. The AUROC for Ctin this cohort was 0.89 (95% CI 0.83 to 0.95, p<0.0001). This study has derived and assessed the diagnostic accuracy of a novel application for 16S PCR. This suggests that 16S PCR in BAL could be used as a rapid test in suspected VAP and may allow better stewardship of antibiotics.
  
  
  
  VAPRAPID trial ref NCT01972425.
authors.affiliation: Division of Anaesthesia, Department of Medicine, University of Cambridge, Cambridge, UK.|Centre for Inflammation Research, University of Edinburgh, Edinburgh, UK.
authors.forename: Andrew
authors.identifier: 0000-0002-3211-3216
authors.initials: A
authors.lastname: Conway Morris
authors.affiliation: Department of Clinical Microbiology, NHS Lothian, Edinburgh, UK.
authors.forename: Naomi
authors.initials: N
authors.lastname: Gadsby
authors.affiliation: Department of Microbiology, Belfast Health & Social Care Trust, Belfast, UK.
authors.forename: James P
authors.initials: JP
authors.lastname: McKenna
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle, UK.
authors.forename: Thomas P
authors.identifier: 0000-0001-5346-7411
authors.initials: TP
authors.lastname: Hellyer
authors.affiliation: Institute of Inflammation and Repair, University of Manchester, Manchester Academic Health Sciences Centre, Manchester, UK.|Intensive Care Unit, Salford Royal NHS Foundation Trust, Greater Manchester, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Dark
authors.affiliation: Intensive Care Unit, Chelsea and Westminster Hospital, Imperial College London, London, UK.
authors.forename: Suveer
authors.initials: S
authors.lastname: Singh
authors.affiliation: Centre for Inflammation Research, University of Edinburgh, Edinburgh, UK.
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Walsh
authors.affiliation: Centre for Infection and Immunity, Queen's University Belfast, UK.|Intensive Care Unit, Royal Victoria Infirmary, Belfast, UK.
authors.forename: Danny F
authors.initials: DF
authors.lastname: McAuley
authors.affiliation: Department of Clinical Microbiology, NHS Lothian, Edinburgh, UK.
authors.forename: Kate
authors.initials: K
authors.lastname: Templeton
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle, UK.
authors.forename: A John
authors.initials: AJ
authors.lastname: Simpson
authors.affiliation: Department of Microbiology, Belfast Health & Social Care Trust, Belfast, UK.|Centre for Infection and Immunity, Queen's University Belfast, UK.
authors.forename: Ronan
authors.initials: R
authors.lastname: McMullan
mesh_terms: D000900:Anti-Bacterial Agents / Q000627:therapeutic use; D015415:Biomarkers / Q000378:metabolism*; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D001999:Bronchoscopy; D015331:Cohort Studies; D006801:Humans; D007362:Intensive Care Units; D053717:Pneumonia, Ventilator-Associated / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics* / Q000382:microbiology; D011237:Predictive Value of Tests; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012680:Sensitivity and Specificity; D006113:United Kingdom
chemical_list: D000900:Anti-Bacterial Agents; D015415:Biomarkers; D012336:RNA, Ribosomal, 16S
keywords: Assisted Ventilation; Bacterial Infection; Pneumonia