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Published on: March 11, 2013
Characterization of cerebrospinal fluid (CSF) microbiota at the time of initial surgical intervention for children
Shailly Pandey1, Kathryn B Whitlock2, Matthew R Test3
1University of Washington School of Medicine, Seattle, Washington, United States of America.
Insights
Bacteria were detected in cerebrospinal fluid (CSF) of children with hydrocephalus using sensitive methods. However, these findings may represent contamination, and the clinical significance of CSF microbiota in these patients remains uncertain.
Area of Science:
- Microbiology
- Pediatric Neurosurgery
- Infectious Diseases
Background:
- Hydrocephalus is a common neurological condition in children.
- The presence and role of microbiota in cerebrospinal fluid (CSF) of pediatric hydrocephalus patients are not well understood.
- Characterizing CSF microbiota is crucial for understanding potential infections and treatment outcomes.
Purpose of the Study:
- To investigate the presence of bacteria in CSF samples from children with hydrocephalus at the time of initial surgery.
- To compare different methods for detecting and characterizing CSF microbiota.
- To assess the potential clinical significance of detected bacteria.
Main Methods:
- CSF samples were collected during initial surgical intervention for hydrocephalus.
- Samples were processed using skim milk-tryptone-glucose-glycerol (STGG) medium storage, standard clinical culture, 16S quantitative polymerase chain reaction (qPCR), and whole-genome amplification sequencing (WGAS).
- Bacterial growth and identification were performed using culture, MALDI-TOF, and sequencing.
Main Results:
- Bacterial growth was observed in 17% of STGG-stored samples and 3% of standard cultures.
- Identified organisms included common skin flora and potential pathogens; only one organism was qPCR positive.
- Whole-genome amplification sequencing (WGAS) and STGG culture showed concordance for only one sample (Staphylococcus epidermidis).
Conclusions:
- Sensitive methods detected bacteria in a subset of pediatric hydrocephalus CSF samples.
- The origin of detected bacteria (true presence vs. contamination) remains uncertain.
- The clinical significance of CSF microbiota in children with hydrocephalus may be limited.
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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