Molecular diagnosis of polymicrobial brain abscesses with 16S-rDNA-based next-generation sequencing

ObjectivesBrain abscesses lead to high mortality despite antibiotic and surgical treatment. Identification of causative bacteria is important guide therapy, but culture-based methods molecular diagnostics by Sanger sequencing 16S PCR products are hampered treatment the often polymicrobial nature brain abscesses. We have applied 16S-rRNA-based next-generation (NGS) for metagenomic analysis intracranial abscess (brain epidural) meningitis samples.MethodsSeventy-nine samples from 54 patients with or were included. DNA was subjected PCR. Amplicons analysed Illumina MiSeq system, sequence reads blasted versus NCBI bacterial database using MEGAN software. Results compared those gram-staining, culture sequencing.ResultsThe NGS workflow successful 51 (46 five nine samples. Inclusion (mono)bacterial allowed us establish a cut-off criterion exclusion contaminating sequences. In total 86 taxa identified in NGS, Streptococcus intermedius Fusobacterium nucleatum as most prevalent species; Propionibacterium Staphylococcus spp. associated epidural two more 31/51 abscesses, revealing these infections allowing discrimination up 16 per sample.ConclusionThese results extend earlier studies showing that expand spectrum detected demonstrate platform suitable this severe infection.