Target identification in Fusobacterium nucleatum by subtractive genomics approach and enrichment analysis of host-pathogen protein-protein interactions

Homology based HP-PPIs predicted from HPIDB. Table S9. Pathway enrichment analysis of host genes from DAVID functional annotation tool. Table S10. Gene ontology report of host genes. Table S11. Disease enrichment analysis of host genes participated in HP-PPIs. Table S12. Total functional annotation cluster analysis of host genes. (XLSX 99 kb

Investigations on microbiome of the used clinical device revealed many uncultivable newer bacterial species associated with persistent chronic infections

Introduction. Chronic persistent device-related infections (DRIs) often give culture-negative results in a microbiological investigation. In such cases, investigations on the device metagenome might have a diagnostic value. Materials and Methods. The 16SrRNA gene sequence analysis and next-generation sequencing (NGS) of clinical metagenome were performed to detect bacterial diversity on invasive medical devices possibly involved in culture-negative DRIs. Device samples were first subjected to microbiological investigation followed by metagenome analysis. Environmental DNA (e-DNA) isolated from device samples was subjected to 16SrRNA gene amplification followed by Sanger sequencing (n=14). In addition, NGS of the device metagenome was also performed (n=12). Five samples were only common in both methods. Results. Microbial growth was observed in only nine cases; among these, five cases were considered significant growth, and in the remaining four cases, growth was considered either insignificant or contaminated. Culture and sequencing analysis yielded identical results only in six cases. In culture-negative cases, Sanger sequencing of 16SrRNA gene and NGS of 16SrDNA microbiome was able to identify the presence of rarely described human pathogens, namely Streptococcus infantis, Gemella haemolysans, Meiothermus silvanus, Schlegelella aquatica, Rothia mucilaginosa, Serratia nematodiphila, and Enterobacter asburiae, along with some known common nosocomial pathogens. Bacterial species such as M. silvanus and S. nematodiphila that are never reported in human infection were also identified. Conclusions. Results of a small number of diverse samples of this pilot study might lead to a path to study a large number of device samples that may validate the diversity witnessed. The study shows that a culture free, a holistic metagenomic approach using NGS could help identify the pathogens in culture-negative chronic DRIs

Additional file 2: Table S4. of Target identification in Fusobacterium nucleatum by subtractive genomics approach and enrichment analysis of host-pathogen protein-protein interactions

Non-human homologous proteins present in F. nucleatum. Table S5. Metabolic pathways of non-human homologous proteins according to KEGG. Table S6. Pathogen-specific metabolic pathways of screened proteins in F. nucleatum. Table S7. Subcellular localization of druggable proteins. (XLSX 45 kb