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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Vydáno v:	APMIS : acta pathologica, microbiologica et immunologica Scandinavica Ročník 127; číslo 10; s. 660 - 670
Hlavní autoři:	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
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	Denmark Wiley Subscription Services, Inc 01.10.2019
Témata:	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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Abstract	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.
AbstractList	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.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. 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. 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.
Author	Jeffries, Thomas T. Biggs, Nathan Radzieta, Michael Bjarnsholt, Thomas Vickery, Karen Dickson, Hugh G. Jensen, Slade O. Fritz, Blaine G. Schwarzer, Saskia Sharma, Varun Malone, Matthew
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BackLink	https://www.ncbi.nlm.nih.gov/pubmed/31344275$$D View this record in MEDLINE/PubMed
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Cites_doi	10.1016/j.cmi.2018.04.036 10.1177/1938640018770285 10.1089/wound.2011.0315 10.1186/s13047-017-0222-5 10.1016/S1473-3099(13)70324-4 10.1371/journal.pone.0009490 10.1086/498112 10.2217/fmb-2017-0174 10.1111/j.1574-695X.2010.00766.x 10.1038/ismej.2012.8 10.2337/dc07-1744 10.1111/iwj.12545 10.1093/nar/gks1219 10.1111/j.1442-9993.1993.tb00438.x 10.1016/j.ebiom.2017.06.026 10.1053/j.jfas.2012.06.017 10.1093/jac/dkx099 10.1086/383271 10.7547/0980290 10.1136/bmjopen-2015-010811 10.1002/dmrr.2699 10.2106/JBJS.17.01152 10.1016/S0140-6736(05)67698-2 10.1053/j.jfas.2010.12.009 10.2337/db18-110-OR 10.3402/dfa.v7.30079 10.1093/cid/cis346 10.1038/nmeth.3869 10.1186/s40168-018-0470-z
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Snippet	Multiple approaches were employed to detect pathogens from bone margins associated with Diabetic Foot Osteomyelitis (DFO). Intra‐operative bone specimens of 14... Multiple approaches were employed to detect pathogens from bone margins associated with Diabetic Foot Osteomyelitis (DFO). Intra-operative bone specimens of 14...
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SubjectTerms	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
Title	Analysis of proximal bone margins in diabetic foot osteomyelitis by conventional culture, DNA sequencing and microscopy
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