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301. The Use of Multiplex Touchdown PCR to Genotype Cutibacterium (Propionibacterium) acnes Isolated from Periprosthetic Shoulder Infections

BACKGROUND: As biogeographic surveys of the human skin microbiome have shown that C. acnes is a major component of the residential axillary microflora, the organism is frequently isolated from synovial tissue and joint aspirates obtained from patients with suspected periprosthetic shoulder infection...

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Autores principales: Nguyen, Frederic, Gorn, Ivan, Desjardins, Marc, Lee, Craig
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6253461/
http://dx.doi.org/10.1093/ofid/ofy210.312
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author Nguyen, Frederic
Gorn, Ivan
Desjardins, Marc
Lee, Craig
author_facet Nguyen, Frederic
Gorn, Ivan
Desjardins, Marc
Lee, Craig
author_sort Nguyen, Frederic
collection PubMed
description BACKGROUND: As biogeographic surveys of the human skin microbiome have shown that C. acnes is a major component of the residential axillary microflora, the organism is frequently isolated from synovial tissue and joint aspirates obtained from patients with suspected periprosthetic shoulder infections. We hypothesized that multilocus sequence typing (MLST) applying a prior validated rapid high through-put multiplex PCR protocol would segregate C. acnes into distinctive phylogroups associated with periprosthetic infections compared with commensal strains. METHODS: C. acnes collected between 2015 and 2017 were correlated with the presence or absence of infection in a detailed retrospective chart review. To determine the C. acnes genotype, bacterial genomic DNA isolated from a single patient isolate served as template in a six locus multiplex touchdown PCR assay using organism-specific primers targeting six genes (16S rRNA, ATPase, sodA, Fic toxin, aspD and recA). Isolates were classified as a contaminant in the absence of multiple positive cultures from an anatomic site and without corresponding clinical, laboratory and histopathologic correlates of infection. The assignment of a diagnosis of prosthetic joint infection (PJI) conformed to the definition recommended by the IDSA Clinical Practice Guidelines of PJI. RESULTS: Of the C. acnes recovered from 94 patients, 14 (14.9%) were from patients with shoulder implants of which shoulder PJI was present in 10 individuals (10.6% of the total). The remaining 84 (89.4%) isolates were retrieved from a variety of tissue and fluid samples of which the majority (65.5%) were deemed as contaminants. Overall, phylogroups IA1, IB, and II predominated (79.8%). Although a similar genetic profile was present in all of the shoulder isolates, no phylogroup association was detected with PJI (P < 0.72). No genetic difference was present in the lineage of strains not causing PJI compared with those responsible for PJI (P < 0.25). CONCLUSION: Our results mirror those from a previous investigation using a less robust four gene MLST PCR based scheme that showed a lack of a phylogenetic association with shoulder PJI. Our results are a reflection of the phylogroup composition of the circulating C. acnes sequence types in our community. DISCLOSURES: All authors: No reported disclosures.
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spelling pubmed-62534612018-11-28 301. The Use of Multiplex Touchdown PCR to Genotype Cutibacterium (Propionibacterium) acnes Isolated from Periprosthetic Shoulder Infections Nguyen, Frederic Gorn, Ivan Desjardins, Marc Lee, Craig Open Forum Infect Dis Abstracts BACKGROUND: As biogeographic surveys of the human skin microbiome have shown that C. acnes is a major component of the residential axillary microflora, the organism is frequently isolated from synovial tissue and joint aspirates obtained from patients with suspected periprosthetic shoulder infections. We hypothesized that multilocus sequence typing (MLST) applying a prior validated rapid high through-put multiplex PCR protocol would segregate C. acnes into distinctive phylogroups associated with periprosthetic infections compared with commensal strains. METHODS: C. acnes collected between 2015 and 2017 were correlated with the presence or absence of infection in a detailed retrospective chart review. To determine the C. acnes genotype, bacterial genomic DNA isolated from a single patient isolate served as template in a six locus multiplex touchdown PCR assay using organism-specific primers targeting six genes (16S rRNA, ATPase, sodA, Fic toxin, aspD and recA). Isolates were classified as a contaminant in the absence of multiple positive cultures from an anatomic site and without corresponding clinical, laboratory and histopathologic correlates of infection. The assignment of a diagnosis of prosthetic joint infection (PJI) conformed to the definition recommended by the IDSA Clinical Practice Guidelines of PJI. RESULTS: Of the C. acnes recovered from 94 patients, 14 (14.9%) were from patients with shoulder implants of which shoulder PJI was present in 10 individuals (10.6% of the total). The remaining 84 (89.4%) isolates were retrieved from a variety of tissue and fluid samples of which the majority (65.5%) were deemed as contaminants. Overall, phylogroups IA1, IB, and II predominated (79.8%). Although a similar genetic profile was present in all of the shoulder isolates, no phylogroup association was detected with PJI (P < 0.72). No genetic difference was present in the lineage of strains not causing PJI compared with those responsible for PJI (P < 0.25). CONCLUSION: Our results mirror those from a previous investigation using a less robust four gene MLST PCR based scheme that showed a lack of a phylogenetic association with shoulder PJI. Our results are a reflection of the phylogroup composition of the circulating C. acnes sequence types in our community. DISCLOSURES: All authors: No reported disclosures. Oxford University Press 2018-11-26 /pmc/articles/PMC6253461/ http://dx.doi.org/10.1093/ofid/ofy210.312 Text en © The Author(s) 2018. Published by Oxford University Press on behalf of Infectious Diseases Society of America. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Abstracts
Nguyen, Frederic
Gorn, Ivan
Desjardins, Marc
Lee, Craig
301. The Use of Multiplex Touchdown PCR to Genotype Cutibacterium (Propionibacterium) acnes Isolated from Periprosthetic Shoulder Infections
title 301. The Use of Multiplex Touchdown PCR to Genotype Cutibacterium (Propionibacterium) acnes Isolated from Periprosthetic Shoulder Infections
title_full 301. The Use of Multiplex Touchdown PCR to Genotype Cutibacterium (Propionibacterium) acnes Isolated from Periprosthetic Shoulder Infections
title_fullStr 301. The Use of Multiplex Touchdown PCR to Genotype Cutibacterium (Propionibacterium) acnes Isolated from Periprosthetic Shoulder Infections
title_full_unstemmed 301. The Use of Multiplex Touchdown PCR to Genotype Cutibacterium (Propionibacterium) acnes Isolated from Periprosthetic Shoulder Infections
title_short 301. The Use of Multiplex Touchdown PCR to Genotype Cutibacterium (Propionibacterium) acnes Isolated from Periprosthetic Shoulder Infections
title_sort 301. the use of multiplex touchdown pcr to genotype cutibacterium (propionibacterium) acnes isolated from periprosthetic shoulder infections
topic Abstracts
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6253461/
http://dx.doi.org/10.1093/ofid/ofy210.312
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