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Orthopaedic Proceedings
Vol. 105-B, Issue SUPP_17 | Pages 42 - 42
24 Nov 2023
Tessier E d'Epenoux Louise R Lartigue M Guerin F Plouzeau-Jayle C Tandé D Chenouard R Bemer P Corvec S
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Abstract Background

The treatment of bone and joint infections (BJI) involving multi-drug resistant bacteria remains a challenge. MDR Staphylococcus epidermidis (MDRSE) clones, resistant to methicillin, clindamycin, levofloxacin, rifampicin and even linezolid, have been reported worldwide. The interest of delafloxacin (DFX), theoretically active on MRSA, remains to be evaluated with respect to MDRSE.

Purpose

Our objective was to evaluate during a retrospective multicenter study the DFX minimal inhibitory concentrations (MICs) and compare its efficacy between ofloxacin-susceptible and ofloxacin-resistant S. epidermidis clinical strains involved in BJI.


Orthopaedic Proceedings
Vol. 105-B, Issue SUPP_17 | Pages 83 - 83
24 Nov 2023
d'Epenoux Louise R Fayoux E Veziers J Dagnelie M Khamari A Deno B Corvec S
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Background

Although described as a commensal bacterium with low pathogenicity, Cutibacterium acnes involvement has been reported in many clinical entities: infections associated with devices, such as shoulder prosthetic joint infections, osteosynthesis, breast implants or cerebrospinal fluid shunts. Various studies show that C. acnes grows as a biofilm, contributing to its persistence by allowing its escape from the action of the immune system and antibiotics.

Purpose

Our aim was to assess the activity of different active substances (erythromycin, clindamycin, doxycycline and Myrtacine®) on eight different well-characterized C. acnes strains after growth in biofilm mode.


Orthopaedic Proceedings
Vol. 105-B, Issue SUPP_17 | Pages 66 - 66
24 Nov 2023
d'Epenoux Louise R Robert M Caillon H Crenn V Dejoie T Lecomte R Tessier E Corvec S Bemer P
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Background

The diagnosis of periprosthetic joint infection (PJI) remains a challenge in clinical practice and the analysis of synovial fluid (SF) is a useful diagnostic tool. Recently, two synovial biomarkers (leukocyte esterase (LE) strip test, alpha-defensin (AD)) have been introduced into the MSIS (MusculoSkeletal Infection Society) algorithm for the diagnosis of PJI. AD, although promising with high sensitivity and specificity, remains expensive. Calprotectin is another protein released upon activation of articular neutrophils. The determination of calprotectin and joint CRP is feasible in a routine laboratory practice with low cost.

Purpose

Our objective was to evaluate different synovial biomarkers (calprotectin, LE, CRP) for the diagnosis of PJI.


Orthopaedic Proceedings
Vol. 104-B, Issue SUPP_10 | Pages 4 - 4
1 Oct 2022
Dupieux C Dubois A Loiez C Marchandin H Lavigne JP Munier C Chanard E Gazzano V Courboulès C Roux A Tessier E Corvec S Bemer P Laurent F Roussel-Gaillard T
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Aim

Bone and joint infections (BJIs) are serious infections requiring early optimized antimicrobial therapy. BJIs can be polymicrobial or caused by fastidious bacteria, and the patient may have received antibiotics prior to sampling, which may decrease the sensitivity of culture-based diagnosis. Furthermore, culture-based diagnosis can take up to 14 days. Molecular approaches can be useful to overcome these concerns. The BioFire® system performs syndromic multiplex PCR in 1 hour, with only a few minutes of sample preparation. The BioFire® Joint Infection (JI) panel (BF-JI), recently FDA-cleared, detects both Gram-positive (n=15) and Gram-negative bacteria (n=14), Candida, and eight antibiotic resistance genes directly from synovial fluids. The aim of this study was to evaluate its performance in acute JIs in real-life conditions.

Method

BF-JI was performed on synovial fluid from patients with clinical suspicion of acute JI, either septic arthritis or periprosthetic JI, in 6 French centers. The results of BF-JI were compared with the results of culture of synovial fluid and other concomitantly collected osteoarticular samples obtained in routine testing in the clinical microbiology laboratory.


Orthopaedic Proceedings
Vol. 103-B, Issue SUPP_15 | Pages 17 - 17
1 Dec 2021
Alier A Torrens C Bellosillo B Gibert J Pérez-Prieto D Corvec S
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Aim

Cutibacterium acnes (C. acnes) is the most cultured organism implicated in periprosthetic shoulder infections. Nevertheless, the clinical significance of its persistence on the skin surface and in the deep layers during shoulder arthroplasty surgery remains still unknown. The purpose of this study was to know if the C. acnes isolate present in deep tissues at the end of a primary shoulder arthroplasty could be responsible for shoulder arthroplasty infection.

Method

Prospective study including 156 patients undergoing primary shoulder arthroplasty. In all the patients included 5 to 12 tissue samples were obtained and were specifically cultured to detect C. acnes presence. DNA was extracted from the C. acnes colonies selected with the QIAsymphony DSP Virus/Pathogen Midi Kit (Qiagen, Hilden, Germany). Libraries were prepared using Nextera XT kit (Illumina) and sequenced in an Illumina MiSeq sequencer. Sequencing files were pre-processed using The Microbial Genome Atlas pipeline. Samples that failed on QC analysis were discarded for further analysis. Isolate nucleotide distances were calculated using Genome-based distance matrix calculator from the enveomics collection. Comparative genomic analysis was performed between intra- and inter-patients’ isolates. Data analysis was performed using R 3.6.3.


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_14 | Pages 69 - 69
1 Dec 2019
Grossi O Lamberet R Touchais S Corvec S Bemer P
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Aim

Cutibacterium acnes is a significant cause of late-onset spinal implant infection (SII). In addition, usual preoperative prophylactic measures may be insufficient to prevent C. acnes operating site colonisation and infection, as demonstrated for prosthetic shoulder surgery. However, little information is available regarding risk factors for SII due to this microorganism. The aims of this study were to determine the characteristics of and risk factors for C. acnes SII.

Method

we conducted a retrospective unmatched case-control study including all adult patients treated for mono and polymicrobial C. acnes SII during 2010–2015. Controls were randomly selected among patients diagnosed with SII due to other microorganisms during the same period.


Aim

Cutibacterium acnes (CA) is one of the crucial actors in spine instrumentation or shoulder prosthesis. Its population is subdivided into 6 major phylotypes: IA1, IA2, IB, IC, II and III. Recent methods for discriminating subpopulations within CA phylotypes highlight the predominance of SLST types H1 to 6 or K1 to 20 in bone and joint infection (BJI). The impact of their ability to produce a biofilm during the development of the infection (with resistance / tolerance to antibiotics used for treatment) remains little studied.

Method

The purpose of this study was to determine whether the ability to establish a biofilm varied according to the different subtypes of clinical strains of CA previously characterized and involved in BJI (hip, knee and shoulder prosthesis). The BioFilm ring test (BioFilm Control®) method with index determination, called BFI (BioFilm Index) inversely proportional to the level of biofilm production was used (BFI = 0.00 indicates a high production of biofilm versus BFI = 20.00 indicates zero production). The BFI was determined after 3 h (T3) and 6 h (T6) incubation. The strains used came from patients, 5 belonging to the IA1 phylotype (SLST A1 and D1 types) and 4 to different phylotypes (IA2, IB, II and III).


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 32 - 32
1 Dec 2018
Aubin G Corvec S
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Aim

Cutibacterium acnes is involved in chronic/low-grade pathologies such as prosthetic joint infection (PJI) or sarcoidosis. During these diseases, granulomatous structures are frequently observed. In this study, we induced a physiological granulomatous reaction in response to different well-characterized clinical C. acnes isolates in order to investigate the cellular process during the granulomatous formation.

Method

An acne C. acnes ATCC6919 isolate (clonal complex (CC) 18, phylotype IA1), a PJI C. acnes BL clinical isolate (CC36, phylotype IB) and a sarcoidosis C. acnes S8 strain (CC28, phylotype IA2) were included and co-culture with PBMC. Cellular aggregation was followed daily using light microscopy. At various time points of incubation (day 3 and day 7), granuloma structures were processed for microscopic observation, colony forming unit enumeration after Triton ×100 lysis to release the internalized bacteria (bacterial load within the granulomas), as well as for flow cytometric analysis (detection of CD4, CD8 and NK lymphocytes).


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 78 - 78
1 Dec 2017
Takoudju E Guillouzouic A Stanimir K Pecorari F Corvec S
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Aim

Although there are no treatment guidelines for Propionibacterium acnes (PA) bone and joint infections (Corvec et al Acta Orthopedica 2016), these infections can be treated with a combination of fluoroquinolones and rifampicin. Rifampicin resistance have already been reported either in in vitro selected mutants or clinical isolates (Furustrand et al JAC 2013, Anaerobe 2015). Minimal inhibitory concentrations of levofloxacin (LVX) ranging from 0,12 to 0.5mg/L are regularly observed but resistance has not yet been investigated. We investigated the in vitro emergence of LVX resistance and characterized the mutations involved in gyrA gene.

Method

The strain of PA ATCC11827 (MIC LVF = 0.25 mg/L) was used. The frequency of mutation was determined after inoculation of 108 PA on blood agar containing concentrations of 2 to 128 times the MIC incubated for 7 days in anaerobiosis at 35 ° C. The emergence of high-level of resistance was also studied from the low-level mutants after a second exposure. For the resistant mutants, the gyrA and parC genes were sequenced and compared to the PA reference sequences.


Orthopaedic Proceedings
Vol. 98-B, Issue SUPP_23 | Pages 70 - 70
1 Dec 2016
Aubin G Gouin F Lepelletier D Jacqueline C Heymann D Asehnoune K Corvec S
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Aim

Propionibacterium acnes is an emerging pathogen especially in orthopedic implant infection. Interestingly, we previously reported a difference in the distribution of the clades involved in spine versus hip or knee prosthetic infection. To date, no study has previously explored the direct impact and close relationship of P. acnes on bone cells according to their own genetic background. The aim of this study was to investigate this interaction of P. acnes clinical strains involved in spine material infections, arthroplasty infections and acne lesions with bone cells.

Method

From a large collection of 88 P. acnes clinical isolates collected between January 2003 and December 2014, a subset of 11 isolates was studied. Four isolates were recovered from spine infections, two from prosthetic infections (knee and hip), three from acne lesions and two reference strains (ATCC11827 and ATCC6919). Implant-associated infections were confirmed according to Infectious Diseases Society of America guidelines for bone and joint infections. Multi-Locus Sequence Typing (MLST) was carried out on all isolates as described by Lomholt et al. PLoS ONE 2010. Bacterial internalization experiments with MG63 osteosarcoma cells were adapted from Crémet et al. Pathog Dis 2015.


Orthopaedic Proceedings
Vol. 97-B, Issue SUPP_15 | Pages 76 - 76
1 Dec 2015
Aubin G Gouin F Lepelletier D Jacqueline C Ashenoune K Corvec S
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Propionibacterium acnes is an emerging pathogen especially in orthopedic implant infection. Aim of this study was to investigate P. acnes phylogeny and to screen for virulence factors among a large collection of clinical isolates involved in spine material infections, arthroplasty infections and acne lesions.

88 P. acnes clinical isolates were collected between January 2003 and December 2014 at Nantes University Hospital (France). Fifty-eight isolates came from spine infections, 14 from prosthetic infections (knee, hip or shoulder), 14 from acne lesions and two reference strains (ATCC11827 and ATCC6919). Implant associated infections were confirmed using Infectious Diseases Society of America criteria for bone and joint infections. Phylotypes and Multi-Locus Sequence Typing (MLST) was carried out on all isolates as described by Lomholt et al. All isolates were tested by established PCR-based assays for 21 putative virulence factor genes characteristic of P. acnes.

MLST analysis revealed an association between clonal complexes (CCs) and origin of P. acnes isolates (p = 0,027). Regarding CCs distribution between different origins, CC36 and phylotype II P. acnes isolates are more frequently observed in prosthetic joint infections. On the other hand, CC18 (IA) and CC28 (IB) P. acnes isolates are more frequently involved in spine infections and acne lesions.

Among all virulence factors screened, hyaluronate lyase gene was only present in CC36 and phylotype II P acnes isolates. Other virulence factors were present in all isolates, whatever their origin or CC.

Regarding molecular typing results, P. acnes involved in spine infections seem to have a skin origin (same CC as isolates from acne lesion). Interestingly, the origin of prosthetic joint infection isolates seems different and they all carry one more virulence factor.

Hyaluronate lyase (Hyl) is a major surface protein of P. acnes with potential antigenetically variable properties that might be essential for P. acnes virulence. Increased tissue permeability caused by the action of hyaluronidase on the extracellular matrix appears to play a role in wound infections, pneumonia, and other sepsis such as bacteremia and meningitis. It could be also take a prominent part in P. acnes prosthetic joint infection pathogenesis.


Orthopaedic Proceedings
Vol. 97-B, Issue SUPP_15 | Pages 31 - 31
1 Dec 2015
Aubin G Lavigne J Guyomarch B Dina C Gouin F Lepelletier D Corvec S
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Staphylococcus aureus is a leading cause of implant-associated infections (IAI). The aim of this study was to identify bacterial and/or clinical features involved in the pathogenesis of S. aureus IAI.

57 IAI S. aureus and 31 nasal carriage (NC) S. aureus isolates were studied. S. aureus genetic background was obtained by microarray analysis. Multi-Locus Sequence Typing was performed to determine clonal complexes (CC). The ability of S. aureus isolates to produce biofilm was investigated by resazurin and crystal violet methods. Clinical data were retrospectively collected from the patient's medical records.

Fifty-five IAI patients were included. Two of them had two different S. aureus IAI episodes. The median age was 73 years (range: 21–96 years) with 29 women (52.7%). The main diagnosis for arthroplasty was arthrosis (38%). Implants were hip prosthesis (n=35), knee prosthesis (n=18) and osteosynthesis (n=4). Infectious and nasal carriage isolates belonged respectively to 18 and 13 different sequence types (STs) without significant difference. Among IAI isolates, five strains were methicillin resistant. IAI isolates were classified as strong (14%), moderate (42.1%) and weak (43.9%) biofilm producers. For NC isolates, distribution was 12.9%, 25.8% and 61.3% for strong, moderate and weak, respectively. Staphylokinase gene was associated with the occurrence of S. aureus IAI (p<0.001). Patients’ ABO blood group phenotype was associated with IAI S. aureus genetic background (sasG, slpB, lukD and set12/ssl8) (p≤0.01). In vitro, CC8 S. aureus strains produce more biofilm than others (p≤0.0001). Two alleles of bbp gene were significantly associated with CC8 S. aureus strains (p≤0.0001). No specific CC involved in IAI compared to NC S. aureus isolates was revealed.

Our results suggested that occurrence of IAI may depend on patients’ ABO blood group and staphylokinase gene detection. We also observed a strong biofilm producer phenotype in CC8 S. aureus. Further studies are needed to prove whether one bbp gene variant is correlated to this phenotype.

This study was supported by a grant number WS1106649 from Pfizer, France and by the French “Ministère de l'Enseignement Supérieur et de la Recherche”.


Orthopaedic Proceedings
Vol. 96-B, Issue SUPP_11 | Pages 283 - 283
1 Jul 2014
Post V Wahl P Uckay I Zimmerli W Corvec S Loiez C Ochsner P Moriarty F
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Summary

Staphylococcus aureus isolates from Fracture fixation device related infections contained fewer isolates that form a strong biofilm in comparison with isolates from Prosthetic joint infections. Both orthopaedic implant related infection groups possessed fnbB and sdrE more frequently than the non-implant related infection groups.

Introduction

One of the most common pathogen causing musculoskeletal infections is Staphylococcus aureus. The aim was to characterise S. aureus isolated from these infections and to look for differences between the isolates from orthopaedic implant related infections (OIRI) and those in non-implant related infections (NIRI). The OIRI are further differentiated in those associated with fracture fixation (FFI) devices and those found in prosthetic joint infections (PJI).