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. 2023 Jun 21;18(6):e0280682.
doi: 10.1371/journal.pone.0280682. eCollection 2023.

Characterization of cerebrospinal fluid (CSF) microbiota at the time of initial surgical intervention for children with hydrocephalus

Shailly Pandey  1 , Kathryn B Whitlock  2 , Matthew R Test  3 , Paul Hodor  4 , Christopher E Pope  3 , David D Limbrick Jr  5   6 , Patrick J McDonald  7   8 , Jason S Hauptman  4   9   10 , Lucas R Hoffman  3   4   10 , Tamara D Simon  11   12   13 ; Cerebrospinal FLuId MicroBiota in Shunts (CLIMB) Study Group
Affiliations

Characterization of cerebrospinal fluid (CSF) microbiota at the time of initial surgical intervention for children with hydrocephalus

Shailly Pandey et al. PLoS One. .

Abstract

Objective: To characterize the microbiota of the cerebrospinal fluid (CSF) from children with hydrocephalus at the time of initial surgical intervention.

Study design: CSF was obtained at initial surgical intervention. One aliquot was stored in skim milk-tryptone-glucose-glycerol (STGG) medium and the second was unprocessed; both were then stored at -70°C. Bacterial growth for CSF samples stored in STGG were subsequently characterized using aerobic and anaerobic culture on blood agar and MALDI-TOF sequencing. All unprocessed CSF samples underwent 16S quantitative polymerase chain reaction (qPCR) sequencing, and a subset underwent standard clinical microbiological culture. CSF with culture growth (either after storage in STGG or standard clinical) were further analyzed using whole-genome amplification sequencing (WGAS).

Results: 11/66 (17%) samples stored in STGG and 1/36 (3%) that underwent standard clinical microbiological culture demonstrated bacterial growth. Of the organisms present, 8 were common skin flora and 4 were potential pathogens; only 1 was also qPCR positive. WGAS findings and STGG culture findings were concordant for only 1 sample, identifying Staphylococcus epidermidis. No significant difference in time to second surgical intervention was observed between the STGG culture-positive and negative groups.

Conclusion(s): Using high sensitivity methods, we detected the presence of bacteria in a subset of CSF samples at the time of first surgery. Therefore, the true presence of bacteria in CSF of children with hydrocephalus cannot be ruled out, though our findings may suggest these bacteria are contaminants or false positives of the detection methods. Regardless of origin, the detection of microbiota in the CSF of these children may not have any clinical significance.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Flow diagram showing sample selection and analysis.
Flow diagram showing number of patients whose CSF samples underwent storage in skim milk-tryptone-glucose-glycerol storage medium (STGG) and subsequent culture with identification by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF), as well as additional analysis with 16S qualitative polymerase chain reaction (qPCR), whole genome amplification sequencing (WGAS), and standard microbiological culture.
Fig 2
Fig 2. Kaplan-Meier curve showing time to second intervention for all children who underwent permanent procedures at first intervention (CSF shunt, ETV).
Stratified by STGG status: STGG positive shown in red, STGG negative shown in blue. p = 0.81.
See this image and copyright information in PMC

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