title: Use of quantitative 16S rRNA PCR to determine bacterial load does not augment conventional cerebrospinal fluid (CSF) cultures among children undergoing treatment for CSF shunt infection.
abstract: The aim of this study was to develop a quantitative 16S rRNA assay for determination of bacterial nucleic acid load in cerebrospinal fluid (CSF) shunt infection and to compare quantitative 16S rRNA polymerase chain reaction (PCR) findings to those of conventional bacterial culture in patients treated for CSF shunt infection. We developed a quantitative 16S rRNA PCR assay that detected bacterial load across a range of 2.5 × 10(9) down to 2.5 × 10(4) 16S copies/mL CSF under experimental conditions for numerous Gram-positive and Gram-negative organisms. However, when applied to archived CSF samples from 25 shunt infection episodes, correlations between positive bacterial culture and 16S rRNA levels were seen in only half of infections, and 16S rRNA levels dropped precipitously after an initial peak on the first day of sample collection. Bacterial load measured using 16S rRNA PCR does not provide sufficient information beyond bacterial culture to inform CSF shunt infection treatment.
authors: Department of Pediatrics, Division of Hospital Medicine, University of Washington/Seattle Children's Hospital, Seattle, WA, USA. Electronic address: Tamara.Simon@seattlechildrens.org.
authors: Tamara D
authors: TD
authors: Simon
authors: Virus, Molecular Biology and Cell Core, Diabetes and Endocrinology Research Center, University of Washington, Seattle, WA, USA.
authors: Brian
authors: B
authors: Van Yserloo
authors: Department of Pediatrics, University of Utah, Salt Lake City, UT, USA.
authors: Kevin
authors: K
authors: Nelson
authors: Department of Neurosurgery, University of Utah, Salt Lake City, UT, USA.
authors: David
authors: D
authors: Gillespie
authors: Department of Neurosurgery, University of Utah, Salt Lake City, UT, USA.
authors: Randy
authors: R
authors: Jensen
authors: Department of Neurosurgery, University of Utah, Salt Lake City, UT, USA.
authors: James P
authors: JP
authors: McAllister
authors: Department of Neurosurgery, University of Utah, Salt Lake City, UT, USA.
authors: Jay
authors: J
authors: Riva-Cambrin
authors: Department of Pediatrics, University of Utah, Salt Lake City, UT, USA.
authors: Chris
authors: C
authors: Stockmann
authors: Microbiology Laboratory, Primary Children's Medical Center, Salt Lake City, UT, USA; Department of Pathology, University of Utah, Salt Lake City, UT, USA.
authors: Judy A
authors: JA
authors: Daly
authors: Department of Pediatrics, University of Utah, Salt Lake City, UT, USA.
authors: Anne J
authors: AJ
authors: Blaschke
mesh_terms: D000293:Adolescent; D000900:Anti-Bacterial Agents / Q000627:therapeutic use; D001419:Bacteria / Q000145:classification / Q000235:genetics*; D001424:Bacterial Infections / Q000175:diagnosis* / Q000188:drug therapy / Q000382:microbiology*; D058491:Bacterial Load; D002555:Cerebrospinal Fluid / Q000382:microbiology*; D002557:Cerebrospinal Fluid Shunts / Q000009:adverse effects*; D002648:Child; D006801:Humans; D012336:RNA, Ribosomal, 16S; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D000900:Anti-Bacterial Agents; D012336:RNA, Ribosomal, 16S
keywords: Bacterial load; Cerebrospinal; Children; Infection; Shunt
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