title: Broad-range real time PCR and DNA sequencing for the diagnosis of bacterial meningitis.
abstract: Rapid aetiological diagnosis of bacterial meningitis is crucial for the early targeting of antimicrobial and adjuvant therapy. Broad-range polymerase chain reaction (PCR) targeting the 16S rRNA gene allows aetiological diagnosis of bacterial meningitis when applied to cerebrospinal fluid (CSF). We assessed the additional diagnostic effect of applying a novel broad-range real time PCR and subsequent DNA sequencing to culture, microscopy, and broad-range conventional PCR on CSF in patients with suspected bacterial meningitis. Broad-range conventional PCR and broad-range real time PCR with subsequent DNA sequencing were applied to 206 CSF specimens collected consecutively from 203 patients aged 6 d to 86 y. Patients' charts were reviewed for clinical information. 17 pathogens were identified by PCR and DNA sequencing or culture. Three specimens were negative by culture but positive by broad-range real time PCR. Three specimens were positive by culture but negative by broad-range real time PCR. Compared with culture, the sensitivity of broad-range real time PCR was 86%, and the specificity 98%. Conventional PCR resulted in a sensitivity of 64% and specificity of 98%. Broad-range real time PCR was generally comparable to culture of CSF and may be a useful supplement, particularly when antimicrobial therapy has been administered. Broad-range real time PCR was more sensitive than broad-range conventional PCR and microscopy.
authors.affiliation: Department of Infectious Diseases, Research Unit Q, Skejby Hospital, Aarhus University Hospital, Denmark. susannadeutch@dadlnet.dk
authors.forename: Susanna
authors.initials: S
authors.lastname: Deutch
authors.forename: Lisbeth N
authors.initials: LN
authors.lastname: Pedersen
authors.forename: Lone
authors.initials: L
authors.lastname: Pødenphant
authors.forename: Rikke
authors.initials: R
authors.lastname: Olesen
authors.forename: Michael B
authors.initials: MB
authors.lastname: Schmidt
authors.forename: Jens K
authors.initials: JK
authors.lastname: Møller
authors.forename: Lars
authors.initials: L
authors.lastname: Ostergaard
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification*; D002648:Child; D002675:Child, Preschool; D004269:DNA, Bacterial / Q000134:cerebrospinal fluid / Q000235:genetics; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D016920:Meningitis, Bacterial / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000382:microbiology*; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004269:DNA, Bacterial