title: Ribosomal PCR and DNA sequencing for detection and identification of bacteria: experience from 6 years of routine analyses of patient samples.
abstract: The use of broad range PCR and DNA sequencing of bacterial 16S ribosomal RNA genes for routine diagnostics of bacterial infections was evaluated. Here, the results from more than 2600 analyses during a 6-year period (2003-2009) are presented. Almost half of the samples were from joints and bones, and the second most frequent origin of samples was from the central nervous system. Overall, 26% of all samples were positive for bacterial DNA and bacterial identification was obtained in 80% of the PCR-positive samples by subsequent DNA sequencing. Ambiguous species identification was noticed among non-haemolytic streptococci, especially within the mitis group. The data show that ribosomal PCR with subsequent DNA sequencing of the PCR product is a most valuable supplement to culture for identifying bacterial agents of both acute and prolonged infections. However, some bacteria, including non-haemolytic streptococci, may not be precisely identified.
authors.affiliation: Department of Microbiological Surveillance and Research, Statens Serum Institut, Copenhagen, Denmark; Department of Science, Systems and Model, Roskilde University, Roskilde, Denmark; Department of Clinical Microbiology, Odense university Hospital, Odense, Denmark.
authors.forename: Kristine Helander
authors.initials: KH
authors.lastname: Jensen
authors.forename: Rimtas
authors.initials: R
authors.lastname: Dargis
authors.forename: Jens Jørgen
authors.initials: JJ
authors.lastname: Christensen
authors.forename: Michael
authors.initials: M
authors.lastname: Kemp
mesh_terms: D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001424:Bacterial Infections / Q000175:diagnosis / Q000235:genetics / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D012336:RNA, Ribosomal, 16S / Q000235:genetics / Q000302:isolation & purification*; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S
keywords: 16S rRNA; Bacteria; DNA sequencing; PCR; identification