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Performance of three ready-to-use bottles for
grinding orthopaedic samples
Olivier Dauwalder1, Camille Kolenda1, Cécile Eymard1, Marine Jawhar1, Cécile Batailler2, Christophe
Gaillard2, Frédéric Laurent1, Céline Dupieux1
1 Bactériologie, Hospices Civils de Lyon, France
2 Chirurgie orthopédique, Hospices Civils de Lyon, France
Céline DUPIEUX
Bacteriology Department - Institute of Infectious Agents - Croix Rousse Hospital
Rhône-Alpes Reference Center for Complex Osteoarticular Infections
celine.dupieux@chu-lyon.fr
Page 2
INTRODUCTION
▪ ▪ Diagnosis of essential IOA by culture (antibiogram)
▪ Grinding of solid samples required before seeding
▪ Objective: optimise sensitivity / limit contamination
▪ Medium: sampling media allowing direct grinding without handling the sample
▪ OBJECTIVE = to compare the analytical performance of two new ready-to-use vials
• ProbeAx Evolution (PAx ; Axonlab A.G.)
• TISSUtainer (TIt ; Diagante) au flacon Ultra-Turrax® UTX-001 (UTx ; Labelians)
Page 3
SUPPORTS EVALUES
Ultra-Turrax® UTX-001 TISSUtainer ProbeAx Evolution
(UTx ; Labelians) (TIt ; Diagante) (PAx ; Axonlab)
Grinding medium Sterile water quality Quality preservation Quality saline solution
BM 2 mL environment BM, 10 mL BM 5 mL
Grinding method Ceramic block + stainless
Steel beeds x10 Steel beeds x4
steel metal grater
Grinder Ultra-Turrax® (IKA) MM400 (Retsch®) SpinAX (AxonLab A.G.)
Grinding time 1 min / 1 sample 2,5 min / 2-8 samples 1 min / 1 sample
Marking CE (old regulations 98/79/ DM-DIV sterile class A, DM-DIV sterile class A,
CE) CE IVDR CE IVDR
Page 4
EQUIPMENT AND METHODS
▪ Prospective study
▪ 2 orthopaedic surgery units at the Hospices Civils de Lyon (HCL)
▪ 15 patients with strong suspicion of IOA
• Diagnostic samples taken for routine care (RC) in Ultra-Turrax tubes
• Among these samples, 2 taken in triplicate and introduced into the operating room in the 3
tubes
• In the laboratory:
- Ultra-Turrax tubes processed according to the RC flow
- ProbeAx and TISSUtainer bottles processed for the study according to the availability of the
technicians in charge (pre-analytical storage at +4°C if waiting; 1-3 days)
Page 5
INTERPRETATION OF RESULTS
▪ Culture reading: quantification/identification of colonies obtained
▪ Definitions:
• Species responsible for AIO: if found ≥ 2 times on SC or study swabs for potentially contaminating
bacteria and ≥ 1 time for non-classically contaminating bacteria
• Contamination if only 1 positive sample with a species that is not very virulent in flora or the
environment
▪ Comparaison :
• Nombre d’espèces bactériennes retrouvées avec chaque poudrier
• Référence : résultats globaux des pvts en Ultra-Turrax du SC
• Pour chaque espèce retrouvée par ≥ 2 types de poudriers, comparaison semi-quantitative du
nombre de colonies obtenues (catégories > 50 UFC, 26-50, 11-25, 6-10, 1-5, uniquement bouillon)
Page 6
RESULTS
▪ The 3 to 7 UTx vials produced for the SC of the 15 patients were
recovered.
• 7 infections monomicrobial
• 5 infections polymicrobial
• no microorganisms for 3 patients
Page 7
RESULTATS
▪ Pour les 3 patients avec prélèvements stériles
Patient Ultra-Turrax SC TISSUTainer ProbeAx Evo
2 2/2 stériles 2/2 stériles 2/2 stériles
5 2/2 stériles 2/2 stériles 2/2 stériles
1/2 stérile
14 2/2 stériles 2/2 stériles
et 1/2 contaminé *
* 1 colonie Cutibacterium acnes sur SCS14
Page 8
RESULTS
▪ For the 7 monomicrobial infections: detected in 6 out of 7 cases (86%)
in at least 1 of the 2 samples in the study, across all vials compared
(UTx, PAx, TIt).
Pvts SC Ultra-Turrax Additional species in
Patient Microorganisme TISSUTainer ProbeAx
globaux SC the study
3 Finegoldia magna 4/5 1/2 1/2 2/2
4 Staphylococcus aureus 5/5 2/2 1/2 1/2
Corynebacterium striatum (TIt et
7 Pseudomonas aeruginosa 5/5 2/2 2/2 2/2 PAx) ; Proteus mirabilis et
Streptococcus constellatus (PAx)
9 Cutibacterium acnes 5/5 2/2 2/2 2/2
10 Staphylococcus aureus 4/4 2/2 2/2 2/2
13 Cutibacterium acnes 2/5 0/2 0/2 0/2 1 contaminant en TIt et 1 en PAx
15 Cutibacterium acnes 5/5 2/2 2/2 2/2
Page 9
RESULTS
▪ For the 5 polymicrobial infections (16 bacterial species in SC): 14 (87.5%)
detected in UTx bottles, 11 (69%) in PAx and 12 (75%) in TIt.
Pvts SC UltraTurrax
Patient Microorganisme TISSUTainer ProbeAx Additional species in the study
globaux SC
1 Escherichia coli 5/7 2/2 2/2 2/2
1 Staphylococcus aureus 1/7 0/2 0/2 0/2
6 Enterobacter cloacae 5/5 2/2 2/2 2/2
6 Enterococcus faecium 5/5 2/2 2/2 1/2
8 Pasteurella canis 2/3 1/2 2/2 2/2
Neisseria spp en plus
8 Fusobacterium spp 2/3 2/2 2/2 2/2
avec PAx et TIt
8 Parvimonas micra 1/3 0/2 0/2 0/2
11 Proteus mirabilis 5/5 2/2 2/2 2/2
1 contaminant en PAx
11 Staphylococcus aureus 2/6 2/2 0/2 0/2
12 Escherichia coli 5/5 2/2 2/2 2/2
12 Staphylococcus aureus 5/5 2/2 2/2 2/2
Staphylococcus epidermidis
12 Enterococcus casseliflavus 4/5 2/2 2/2 2/2
en plus avec PAx et TIt
12 Acinetobacter nosocomialis 5/5 2/2 1/2 2/2
1 contaminant en PAx
12 Lysinibacillus fusiformis 2/5 1/2 0/2 0/2
12 Bacillus thuringiensis 5/5 2/2 2/2 2/2
12 Streptococcus mitis-oralis 4/5 1/2 2/2 0/2
Page 10
RESULTS - SUMMARY
Overall SC results (3
Results for 2 UTx Results for 2 TIt Results for 2 PAx
to 7 UTx)
3 patients with
stériles stériles stériles
sterile samples
- - 6 detected by ≥ 1 of the - - 6 detected by ≥ 1 of - - 6 detected by ≥ 1 of
7 IOA monomicrobial samples included the samples included the samples included
- - 1 not detected - - 1 not detected - - 1 not detected
5 IOA polymicrobial (16 5/5 detected 5/5 detected 5/5 detected
species) (14 species) (12 species) (11 species)
Additional species:
Additional species :
- contaminations: 3
- contaminations: 2 (C.
(non-aureus staphylococci)
acnes, Kocuria)
- pathogens: 5
- pathogens: 3
(C. striatum, Neisseria spp,
(C. striatum, Neisseria spp,
P. mirabilis, S. epidermidis,
S. epidermidis)
S. constellatus)
→ Ultimately, the results from the new bottles were strictly consistent or detected more
species than the SC UTx in 76.7% of cases for both PAx and TIt.
Page 11
RESULTS
▪ Pre-analytical storage at +4°C for up to 3 days of
PAx and TIt bottles did not prevent the detection of
fastidious species (Pasteurella, Neisseria,
anaerobes).
▪ The number of colonies obtained on the culture
media was not significantly different despite
different grinding volumes depending on the flasks
(UTx: 2 mL; PAx: 5 mL; TIt: 10 mL).
Semi-quantitative categorization: > 50 CFU,
26-50, 11-25, 6-10, 1-5, broth only
Page 12
DISCUSSION-CONCLUSION
▪ Prospective study, samples introduced into the new powder containers as soon as
they leave the operating room
▪ small number of patients -> larger prospective studies needed
▪ Demonstrates at least equivalent analytical performance between the new TISSUtainer
and ProbeAx Evo vials vs. Ultra-Turrax.
▪ Other practical criteria should be taken into account. :
• bag and bottle holder,
• impact resistance (pneumatic),
• noise (UTx > TIt > PAx),
• space requirement (TIt > UTx et PAx),
• number of samples processed at the same time (TIt > Utx et PAx)
• or volume of available mulch (TIt > PAx > UTx)
Page 13
ACKNOWLEDGEMENTS
▪ To the technicians and biologists of the bacteriology department
at the Institute of Infectious Agents, Hospices Civils de Lyon
Cécile EYMARD Olivier DAUWALDER
Marine JAWHAR Camille KOLENDA
▪ To orthopedic surgeons who included patients
Pr Cécile BATAILLER
Dr Christophe GAILLARD