title: Comparison between a Broad-Range Real-Time and a Broad-Range End-Point PCR Assays for the Detection of Bacterial 16S rRNA in Clinical Samples.
abstract: Broad range PCR targeting the 16S rRNA gene is widely used to test clinical samples for the presence of bacterial DNA. End-point 16S PCR is both time-consuming and at high risk of cross-contamination. Prior to the replacement of the 16S end-point PCR assay routinely used in our clinical laboratory by a new 16S real-time PCR assay, we aimed to compare the performances of both techniques for the direct diagnosis of bacterial infections in clinical samples. In this prospective study, 129 clinical samples were included for direct comparison of both techniques. The sensitivity of 16S real-time PCR assay (76%) was significantly higher than that of end-point 16S PCR assay (41%) (p<0.01). Specificities of both PCR assays did not differ significantly (p=0.43). The 16S real-time PCR assay yielded an etiological diagnosis in 19% of culture-negative samples. It constitutes a reliable and complementary diagnostic tool to the bacterial culture.
authors.affiliation: Faculty of Medicine, University of Strasbourg, Strasbourg, France.
authors.forename: Mariam
authors.initials: M
authors.lastname: Meddeb
authors.affiliation: Faculty of Medicine, University of Strasbourg, Strasbourg, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Koebel
authors.affiliation: Faculty of Medicine, University of Strasbourg, Strasbourg, France.
authors.forename: Benoît
authors.initials: B
authors.lastname: Jaulhac
authors.affiliation: Faculty of Medicine, University of Strasbourg, Strasbourg, France frederic.schramm@unistra.fr.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Schramm
mesh_terms: D001424:Bacterial Infections / Q000175:diagnosis; D023381:Endpoint Determination; D006801:Humans; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards
chemical_list: D012336:RNA, Ribosomal, 16S
keywords: 16S rRNA; broad-range PCR; molecular diagnosis; real-time PCR