title: Diagnostic utility of broad range bacterial 16S rRNA gene PCR with degradation of human and free bacterial DNA in bloodstream infection is more sensitive than an in-house developed PCR without degradation of human and free bacterial DNA.
abstract: We compared a commercial broad range 16S rRNA gene PCR assay (SepsiTest) to an in-house developed assay (IHP). We assessed whether CD64 index, a biomarker of bacterial infection, can be used to exclude patients with a low probability of systemic bacterial infection. From January to March 2010, 23 patients with suspected sepsis were enrolled. CD64 index, procalcitonin, and C-reactive protein were measured on admission. Broad range 16S rRNA gene PCR was performed from whole blood (SepsiTest) or blood plasma (IHP) and compared to blood culture results. Blood samples spiked with Staphylococcus aureus were used to assess sensitivity of the molecular assays in vitro. CD64 index was lower in patients where possible sepsis was excluded than in patients with microbiologically confirmed sepsis (P = 0.004). SepsiTest identified more relevant pathogens than blood cultures (P = 0.008); in three patients (13%) results from blood culture and SepsiTest were congruent, whereas in four cases (17.4%) relevant pathogens were detected by SepsiTest only. In vitro spiking experiments suggested equal sensitivity of SepsiTest and IHP. A diagnostic algorithm using CD64 index as a decision maker to perform SepsiTest shows improved detection of pathogens in patients with suspected blood stream infection and may enable earlier targeted antibiotic therapy.
authors.affiliation: Novo Mesto General Hospital, 8000 Novo Mesto, Slovenia.
authors.forename: Petra
authors.initials: P
authors.lastname: Rogina
authors.affiliation: Institute of Microbiology and Immunology, Faculty of Medicine, 1000 Ljubljana, Slovenia.
authors.forename: Miha
authors.identifier: 0000-0003-0170-1733
authors.initials: M
authors.lastname: Skvarc
authors.affiliation: Institute of Microbiology and Immunology, Faculty of Medicine, 1000 Ljubljana, Slovenia.
authors.forename: David
authors.identifier: 0000-0002-9653-9830
authors.initials: D
authors.lastname: Stubljar
authors.affiliation: Institute of Microbiology and Immunology, Faculty of Medicine, 1000 Ljubljana, Slovenia.
authors.forename: Romina
authors.initials: R
authors.lastname: Kofol
authors.affiliation: Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne 50935, Germany.
authors.forename: Achim
authors.initials: A
authors.lastname: Kaasch
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D016470:Bacteremia / Q000097:blood* / Q000175:diagnosis*; D001424:Bacterial Infections / Q000097:blood* / Q000175:diagnosis*; D004269:DNA, Bacterial / Q000032:analysis* / Q000097:blood / Q000235:genetics*; D006801:Humans; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000032:analysis* / Q000235:genetics*; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D055815:Young Adult
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S