All methods were performed using manufacturers’ suggested protocols. Following

sequencing the individual sequence reads were screened to provide a final library of quality trimmed reads > 200 bp. These reads were then analyzed using IMG/M Expert Review metagenomics analysis system of the joint genome institute http://​www.​jgi.​doe.​gov. Individual reads were not QNZ research buy assembled prior to analysis and only reads providing hits based upon IMG/M criteria [34, 35] were utilized in the analyses. Due to HIPAA issues this data is not publically available but the microbial data has been deconvoluted and submitted to the SRA as indicated below. Quantitative PCR Using a quantitative PCR wound diagnostic panel (Pathogenius diagnostics, Lubbock, TX), described previously [12, 16] 8 of the learn more 40 VLU samples, chosen because they contained a predicted predominance of bacteria targeted by the qPCR wound panel were evaluated. The

results of the qPCR were provided in the form of relative ratios of each detected bacteria in the sample and these results compared to corresponding bTEFAP bacterial ratio data. Data submission and availability at NCBI The data from the bTEFAP analyses and microbial metagenomic data are available in the National Center for Biotechnology Information’ http://​www.​ncbi.​nlm.​nih.​gov short read archive (SRA) under project accession number [GenBank:SRA008389.2/VLU]. Basic Statistics Statistics were performed using comparative functions and multivariate hierarchical clustering methods PRKACG of NCSS 2007 (NCSS, Kaysville, Utah). Acknowledgements Written consent for publication was obtained from the patient or their relative. The letter of informed consent utilized for this study in relation to Western Institutional Review Board Protocol # 20062347 has been provided to BMC microbiology editors. Signed consent forms for study participation and photos are on file at Southwest Regional Wound Care Center (Lubbock, TX) for all patients participating in this IRB approved study. This research was funded by internal research monies of MBRI, which is directly toward elucidation

of microbial diversity associated with chronic infections and biofilms. Southwest Regional Wound Care Center is dedicated to improving patient care and outcomes in relation to chronic wounds and has deemed that this website scientific publications are the fastest method to distributing knowledge to help patients and clinicians dealing with chronic wounds worldwide. Research and Testing Laboratory is a for-profit service laboratory providing molecular testing and bTEFAP analysis to the public. Electronic supplementary material Additional file 1: Spreadsheet of bacterial genera detected among VLU. The data file provides a complete compiled output of the bacterial genera detected among the VLU. (XLS 32 KB) Additional file 2: Spreadsheet of VLU topology for subjects 5, 6, 7, and 8.