Analysis of proximal bone margins in diabetic foot osteomyelitis by conventional culture, DNA sequencing and microscopy

Multiple approaches were employed to detect pathogens from bone margins associated with Diabetic Foot Osteomyelitis (DFO). Intra‐operative bone specimens of 14 consecutive subjects with suspected DFO were collected over a six‐month study period from Liverpool Hospital. Infected bone and a proximal b...

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Bibliographic Details
Published in:APMIS : acta pathologica, microbiologica et immunologica Scandinavica Vol. 127; no. 10; pp. 660 - 670
Main Authors: Malone, Matthew, Fritz, Blaine G., Vickery, Karen, Schwarzer, Saskia, Sharma, Varun, Biggs, Nathan, Radzieta, Michael, Jeffries, Thomas T., Dickson, Hugh G., Jensen, Slade O., Bjarnsholt, Thomas
Format: Journal Article
Language:English
Published: Denmark Wiley Subscription Services, Inc 01.10.2019
Subjects:
16S rRNA
Bacteria - classification
Bacteria - genetics
Bacteria - isolation & purification
Bacteriological Techniques - methods
Biomedical materials
Bone and Bones - microbiology
Bone and Bones - surgery
Bone growth
Culture
Deoxyribonucleic acid
Diabetes
Diabetes mellitus
diabetic foot
Diabetic Foot - microbiology
Diabetic Foot - pathology
Diabetic Foot - surgery
DNA
DNA sequencing
Feet
Foot diseases
Histocytochemistry - methods
Humans
In Situ Hybridization, Fluorescence
Metagenomics - methods
Microbiomes
Microorganisms
Microscopy
Microscopy, Electron, Scanning
Osteomyelitis
Osteomyelitis - microbiology
Osteomyelitis - pathology
Osteomyelitis - surgery
Pathogens
Sequence Analysis, DNA
Statistical analysis
surgical management
Osteomyelitis
16S rRNA
surgical management
diabetic foot
ISSN:0903-4641, 1600-0463, 1600-0463
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Summary:Multiple approaches were employed to detect pathogens from bone margins associated with Diabetic Foot Osteomyelitis (DFO). Intra‐operative bone specimens of 14 consecutive subjects with suspected DFO were collected over a six‐month study period from Liverpool Hospital. Infected bone and a proximal bone margins presumed to be ‘clean/non‐infected’ were collected. Bone material was subjected to conventional culture, DNA sequencing and microscopy. In total, eight of 14 (57%) proximal bone margins had no growth by conventional culture but were identified in all proximal bone specimens by DNA sequencing. Proximal margins had lower median total microbial counts than infected specimens, but these differences were not statistically significant. Pathogens identified by sequencing in infected specimens were identified in proximal margins and the microbiomes were similar (ANOSIM = 0.02, p = 0.59). Using a combination of SEM and/or PNA‐FISH, we visualized the presence of microorganisms in infected bone specimens and their corresponding proximal margins of seven patients (50%) with DFO. We identify that bacteria can still reside in what seems to be proximal ‘clean’ margins. The significance and implications of clinical outcomes requires further analysis from a larger sample size that incorporates differences in surgical and post‐operative approaches, correlating any outcomes back to culture‐sequence findings.
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ISSN:0903-4641
1600-0463
1600-0463
DOI:10.1111/apm.12986