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Does intraoperative tissue sample enrichment help or hinder the identification of microorganisms in prosthetic joint infection?


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Objectives Biofilm formation reduces the efficacy of standard microbiological techniques in prosthetic joint infection. This study aimed to investigate the sensitivity, specificity and predictive values of tissue sample enrichment as a means to increase diagnostic yield. Methods Patients undergoing revision arthroplasty surgery between May 2004 and January 2013 had intraoperative tissue samples cultured in standard media as well as enriched in brain heart infusion broth. Patients were separated into infected or non-infected groups according to modified criteria from the Musculoskeletal Infection Society. Results A total of 197 revision arthroplasties were included (non-infected, n = 165; proven infection, n = 32). The mean time until revision in non-infected and infected groups was 75.9 and 41.7 months, respectively. The commonest microorganisms cultured were coagulase-negative staphylococci (42.9 %) and Staphylococcus aureus (34.4 %). The sensitivity and specificity of standard tissue culture were 0.25 (CI 0.18–0.33) and 0.98 (CI 0.95–0.99), respectively. Including enrichment culture results increased the sensitivity to 0.45 (CI 0.37–0.54), but decreased specificity to 0.59 (CI 0.52–0.66). Conclusion Any potential increase in the sensitivity is far outweighed by the extremely high false-positive rate. Results of tissue samples cultured by enrichment should be used with caution and may lead to a worse outcome if incorrectly interpreted.
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Authors pre-print manuscript. Published version available at Eur J Orthop Surg Traumatol. 1
2014 Nov 22. [Epub ahead of print] PMID: 25416208 2
Does intraoperative tissue sample enrichment help or hinder the 4
identification of microorganisms in prosthetic joint infection? 5
Running title: 8
Intra-operative tissue sample enrichment for diagnosis of prosthetic joint infection 9
Mr Robert W Jordan1, Specialist Registrar 13
Mr Adnan Saithna2*, Consultant Orthopaedic Surgeon 15
E-mail: 16
Tel: +44(0)247 6351 351 17
Mr Nicholas Smith1, Specialist Registrar 19
Mr Rory Norris1, Specialist Registrar 21
Mr Andrew Sprowson3, Associate Professor 23
Mr Pedro Foguet1, Consultant Orthopaedic Surgeon 25
*Denotes corresponding author 27
1. University Hospitals Coventry and Warwickshire NHS Trust, Clifford Bridge Road, 31
Walsgrave, Coventry, CV2 2DX, UK 32
2. The George Eliot Hospital, College Street, Nuneaton, CV10 7DJ, UK 34
3. The University of Warwick, Coventry, CV4 7AL, UK 36
Abstract 45
Objectives: Biofilm formation reduces the efficacy of standard microbiological techniques in 46
prosthetic joint infection. This study aimed to investigate the sensitivity, specificity and 47
predictive values of tissue sample enrichment as a means to increase diagnostic yield. 48
Methods: Patients undergoing revision arthroplasty surgery between May 2004 and January 50
2013 had intra-operative tissue samples cultured in standard media as well as enriched in 51
Brain Heart Infusion Broth. Patients were separated into infected or non-infected groups 52
according to modified criteria from the Musculoskeletal Infection Society. 53
Results: 197 revision arthroplasties were included (non infected, n=165; proven infection, 55
n=32). The mean time until revision in non-infected and infected groups was 75.9 and 41.7 56
months respectively. The commonest microorganisms cultured were coagulase negative 57
staphylococci (42.9%) and staphylococcus aureus (34.4%). The sensitivity and specificity of 58
standard tissue culture was 0.25 (CI 0.18-0.33) and 0.98 (CI 0.95-0.99). Including enrichment 59
culture results increased the sensitivity to 0.45 (CI 0.37-0.54) but decreased specificity to 60
0.59 (CI 0.52-0.66). 61
Conclusion: Any potential increase in the sensitivity is far outweighed by the extremely high 63
false positive rate. Results of tissue samples cultured by enrichment should be used with 64
caution and may lead to a worse outcome if incorrectly interpreted. 65
Key words: 67
Arthroplasty; Prosthetic Joint Infection; Revision Surgery; Microbiology; Tissue Enrichment 68
Culture 69
Introduction 70
Infection after arthroplasty surgery is a devastating complication with an incidence of around 71
1%.1-4 Diagnosis can be challenging due to the formation of biofilms, the resulting low 72
diagnostic yield of standard microbiological techniques and the lack of consensus for 73
defining infection. In order to address this, the Musculoskeletal Infection Society has recently 74
published criteria for prosthetic joint infection (PJI) that uses a combination of clinical, 75
microbiological, biochemical and histological results.5 76
Correct identification of the causative microorganism is essential to optimize clinical 77
outcomes as this allows appropriately targeted antibiotic therapy based on culture 78
sensitivities. The culture of intraoperative tissue samples has traditionally been the gold 79
standard for identifying microorganisms in PJIs.6,7 Five or six tissue samples should be 80
harvested and a positive result is reflected by 3 or more positive cultures.8 However, the 81
sensitivity of these tests is often low in PJI due to biofilm formation.8-10 This has led to the 82
exploration of further techniques to increase the sensitivity of microorganism detection, such 83
as culture enrichment. This process involves placing the sample in a ‘broth’, which provides 84
ideal conditions for growth of microorganisms. Studies have reported that enrichment can 85
increase the sensitivity of both generic samples11 and within a PJI setting.12, 13 However some 86
authors argue that the increase in positive results is in fact a result of sample contamination.14 87
Therefore the exact role of enrichment in helping identify microorganisms in PJI remains 88
unclear and this study aims to assess the role of enrichment in terms of sensitivity, specificity, 89
and predictive values. 90
Methods 92
We retrospectively reviewed all revision arthroplasty procedures performed by the senior 93
surgeon (PF) for any indication between May 2004 and January 2013. During this period 94
both one and two stage revision surgeries were undertaken. Those procedures performed as a 95
one-stage revision or the first stage of a two-stage revision were included for analysis in this 96
study. The Musculoskeletal Infection Society published criteria for diagnosis of PJI in 2011, 97
however a modification of this criteria was used to classify patients as either infected or non-98
infected post-operatively (see Table 1).5 This modification was necessary as during the study 99
period as neither histology nor synovial white blood cell counts were performed on intra-100
operative tissue samples at our centre. It is our understanding that this reflects practice in 101
many centres and therefore represents a pragmatic study design. 102
Table 1 - Criteria used for diagnosis of peri-prosthetic joint infection 104
1)# Sinus#tract#communicating#with#the#prosthesis.
2)## Pathogen#isolated#by#culture#from#2#or#more#####
1)# Elevated#erythrocyte#sedimentation#rate#and#
2)# ‘Profuse’#or#‘high’#synovial#white#blood#cell##
3)# ‘Majority’#of#polymorphonuclear#celles#in####
4)# Presence#of#purulence#in#the#affect#joint.
5)# Isolation#of#a#microorganism#in#one#culture#of##
Revision surgery was performed by the senior author (PF) in a theatre with laminar airflow. 106
A minimum of 3 different tissue samples were taken in all infected cases, taken with a fresh 107
sterile scalpel and forceps and put into separate containers. Tissue samples were taken in an 108
identical fashion for non-infected revisions but cases where no tissue samples were taken 109
were excluded from the study. On arrival in the laboratory tissue samples were divided into 110
two; the first part was cultured on standard agar plates for a total of 5 days and the second 111
part underwent enrichment. Results from the two culturing processes were reported 112
separately; one for standard culture and the other for enrichment. The enrichment process 113
involved placement of the sample in a Brain Heart Infusion Broth (Oxoid Microbiology 114
Products, Hampshire, UK).15 After 24 hours in the enrichment broth the sample was then 115
placed on standard agar plates for a further five days. 116
For those in the infected group empirical intravenous antibiotics were commenced post-118
operatively until microbiological results allowed tailoring of the treatment strategy. Once the 119
patients C-reactive protein had normalised they were converted to an oral antibiotic regimen 120
whilst an inpatient. If after 48 hours of oral antibiotics the patient remained well and the C-121
reactive protein normal they could be discharged with outpatient appointments at 1, 3 and 6 122
weeks after which antibiotics were stopped if progress was satisfactory. 123
Hospital electronic records were used to retrieve data including demographics, primary 125
diagnosis, date of surgery, reason for failure of arthroplasty, pathology and microbiology 126
results, clinic letters, theatre records, the use of antibiotics prior to revision surgery and the 127
requirement for further surgery. Microbiological results from all tissue cultures were 128
recorded, assessing both the number of specimens taken and the number of positive results by 129
culture and enrichment. Isolation of a microorganism in the infected group was taken as a 130
true positive result and in the non-infected as a false positive. A negative culture was 131
regarded as a false negative in the infected group and a true negative in the non-infected 132
group. Patients were followed up after their revision surgery until July 2013 to analyse 133
whether further surgery was required and to determine whether there was any evidence of 134
infection. When further surgery had been undertaken, both the indication for surgery and the 135
microbiological results from tissue samples taken were collected. Sensitivity, specificity and 136
positive and negative predictive values, with their respective binomial 95% confidence 137
intervals (CIs), were calculated by using standard formulae. 138
Results 140
217 revisions were performed during the study period. Using the modified criteria 32 were 141
classified as infected (16 hips and 16 knees), and 185 cases as non-infected. All 32 infected 142
cases had at least 3 tissue samples taken but 20 of those in the non-infected group had no 143
tissue samples taken and were excluded from the study. After exclusion, a total of 197 cases 144
were included in the study, of which 165 were non-infected (53 knee and 112 hip revisions) 145
and 32 were infected. The demographics of the two groups are shown in Table 2. 146
Table 2 - Patient demographics 156
Non-infected group
Infected group
Age (years)
Gender (%)
Primary diagnosis
Post fracture
In the infected group, the mean age was 69.5 years and 56.3% were female. 24 were 158
performed as a one-stage revision and 8 were the first stage of a two-stage revision. The 159
mean time from original surgery to revision was 41.7 months (range 7 to 148 months) and 160
this is further illustrated in Table 3. A total of 141 tissue samples were collected with 35 161
positive on standard cultures and 29 by enrichment. The commonest microorganisms grown 162
in standard cultures were coagulase negative staphylococci (42.9%) and staphylococcus 163
aureus (34.4%). The microorganisms isolated in all samples are shown in Table 4. Twelve 164
patients had received oral antibiotics within six months of their revision surgery and in these 165
cases 49 tissue samples were harvested of which 6 (10%) were positive on standard cultures 166
and 11 (27%) by enrichment, further details are shown in Table 5. 167
Table 3 – Stratification of patients by time elapsed from original surgery until revision 170
Non-infected group (%)
Infected group (%)
Under 1 year
1 to 2 years
2 to 3 years
3 to 4 years
4 to 5 years
5 to 10 years
10 to 15 years
15 to 20 years
Over 20 years
Table 4 - Microorganisms isolated from tissue culture in infected cases 173
Positive standard culture
Positive enrichment culture
Coagulase negative
Staphylococcus aureus
Enterobacter cloacae
Alpha haemolytic
Table 5 - Effect of pre-operative antibiotics on standard tissue culture results 176
Number of
Number of
samples (%)
Until surgery
1 month prior to
1-3 months
prior to surgery
3-6 months
prior to surgery
Any antibiotics
within 6 months
No antibiotics
The non-infected group contained 165 patients with a mean age of 63.9 years. The mean time 178
until revision was 75.9 months (range 6 months to 434 months) as detailed in Table 3. The 179
indications for revision surgery are illustrated in Table 6 with the commonest being aseptic 180
loosening (44.8%), problematic resurfacing (21.1%) and recurrent dislocations (9.7%). In this 181
group a total of 215 tissue samples were taken with 4 positive in standard cultures and 84 by 182
enrichment. The microorganisms grown are shown in Table 7 with coagulase negative 183
staphylococcus (72.6%) being isolated in the majority of positive enrichment samples 184
Table 6 - Reason for prosthesis failure in the non-infected group 188
Aseptic loosening
Painful hip resurfacing
Pain with no obvious cause
Progression of arthritis to other knee
Failed cannulated screw
Pain post hemiarthroplasty
Impending fracture
The sensitivity and specificity of standard tissue culture was 0.25 (CI 0.18-0.33) and 0.98 (CI 190
0.95-0.99). Including data from enrichment studies increased the sensitivity to 0.45 (CI 0.37-191
0.54) but decreased specificity to 0.59 (CI 0.52-0.66). Similarly, the positive predictive value 192
decreased from 0.90 (CI 0.75-0.97) to 0.42 (CI 0.34-0.50). The negative predictive value was 193
virtually unchanged, from 0.67 (CI 0.61-0.77) to 0.62 (CI 0.55-0.69). These results are shown 194
in Table 8 and similar findings were demonstrated in both hip and knee cases. 195
Patients were followed up after revision surgery for a mean of 45.7 months in the infected 197
group (range 6 to 105) and 40.8 months in the non-infected group (range 6 to 108.7). During 198
this follow up period, 7 of the 32 infected cases required further surgery with the details 199
shown in Table 9. Of these, four required further surgery for infection. In the non-infected 200
group 19 (11.5%) required further surgery during follow up; 10 for recurrent dislocations, 5 201
for aseptic loosening and 4 for infection. The mean time to further surgery in the four 202
infected cases was 2.7 months, in three of these cases more than 2 tissue cultures were 203
positive and in the other all samples were negative. None of these four patients had grown 204
any organism by either enrichment or standard culture at their initial revision surgery. 205
Table 7 - Microorganisms isolated from tissue culture in non-infected cases 207
Positive standard culture
Positive enrichment
Coagulase negative
Staphylococcus aureus
Skin flora
Alpha haemolytic
Streptococcus mitis
Escherichia coli
Bacillus oxidase
Table 8 - Sensitivity, specificity, positive and negative predictive values 209
All Tissue
samples (CI)
All Tissue
samples +
TKR tissue
samples (CI)
TKR tissue +
samples (CI)
THR tissue
samples (CI)
THR tissue +
samples (CI)
Table 9 - Further surgery during follow up of infected cases 215
Original revision
Reason for
further surgery
One stage hip
Required revision surgery – intraoperative
samples negative
Required revision surgery – intraoperative
samples negative
3 months post procedure - same organism grown
as original procedure. Treated with suppression
Two stage hip
One stage knee
Patella tendon
Required reconstructive surgery – intraoperative
samples negative
Two stage knee
Nine months post surgery - grew streptococcus,
previous samples negative.
Treated with debridement, change of spacer and
antibiotics successfully.
44 months post surgery - grew pseudomonas,
same organisms as initial surgery
28 months post surgery - grew pseudomonas,
previous samples negative. Treated by above
knee amputation after failed debridements.
Discussion 218
Biofilm formation in PJI reduces the diagnostic yield of standard microbiological techniques. 219
Enrichment has been used to try and increase the detection rate as identifying the causative 220
organism allows targeted antibiotic therapy and improved outcomes.11-13,16 However in our 221
study 39% of samples taken in non-infected cases were positive by enrichment and this 222
supports concerns that sample contamination is responsible for the increased number of 223
microorganisms isolated and an unacceptable increase in the rate of false positive results.14 224
The primary goal of identifying the responsible microorganism intra-operatively is to allow 226
targeted antibiotic treatment.16 If a microorganism is not identified, the patient is usually 227
treated with broad spectrum antibiotics. If a microorganism is detected, targeted antibiotics 228
can be used. However if the incorrect microorganism is identified, targeted antibiotics may 229
not cover the ‘true’ microorganism. Therefore in cases of already known infection, the 230
specificity can be considered more important. Inclusion of results from enrichment reduced 231
the specificity in our study from 0.98 to 0.59 with a similar reduction reported in both THR 232
and TKR cases. This suggests using enrichment results would increase the number of patients 233
receiving a false positive diagnosis (and unnecessary antibiotic treatment) and also increase 234
the risk of prescribing incorrectly targeted antibiotics in cases of true infection. 235
The commonest microorganisms cultured in our study were coagulase negative 237
staphylococcus and staphylococcus aureus and this reflects their known incidence in PJI.17-20 238
In those patients receiving antibiotics within 6 months of revision surgery the proportion of 239
positive standard tissue cultures was 10.2% compared to 32.6% in the those patients who did 240
not receive antibiotics. This is consistent with previous work which demonstrated that over 241
50% of culture negative PJIs follow treatment with antibiotics.21 As, there is no single test 242
that can diagnose PJI, it is possible that some of the non-infected group were suffering from 243
low grade infections that were not identified at the time of revision surgery. We followed up 244
these patients for a mean of 75.9 months to assess whether they had required subsequent 245
surgery and if there was any evidence of on-going infection. During this follow up period 246
four patients (2.4%) required further surgical intervention for infection with three having 247
positive microbiological results at the time of this surgery. Interestingly, none had previously 248
grown organisms by either standard culture or enrichment at their original revision surgery 249
suggesting that these infections were either de novo from the time of revision surgery or were 250
unidentified infections. Previous studies have reported infection rates in aseptic revisions of 251
between 1.3% and 5% and the fact our results are similar would suggest that these may also 252
be de novo infections.22-24 253
As well as the difficulty in definitively diagnosing or excluding PJI, there are some 255
limitations to this study. The retrospective design resulted in some slight variability in 256
practice. There were a total of 20 non-infected cases where tissue samples were not harvested 257
and in the remainder of this group the numbers of samples varied, which may bias the results. 258
A positive culture in the infected group for both standard and enrichment cultures was 259
assumed to be a true positive and this is likely to have over-estimated the sensitivity of these 260
techniques. We know from the non-infected group that enrichment produced a high false 261
positive rate and therefore a proportion of positive cultures in the infected group may also 262
have been secondary to contamination. In addition, we only analysed tissue sample cultures. 263
Therefore the results of this study are only valid when considering tissue samples intra-264
operatively. 265
Conclusion 268
This study has produced some important results that can help clinicians determine whether 269
the use of BHIB enrichment cultures are indicated in the investigation of a particular patient 270
and how to interpret the results. We conclude that there is an unacceptably high false 271
positive rate in presumed non-infected cases and great caution should be taken in diagnosing 272
infection in these patients on the basis of BHIB enrichment cultures alone. In those patients 273
with a clinical PJI, standard microbiological techniques have low yield and using BHIB 274
enrichment cultures can increase sensitivity. However false positive results are still a concern 275
and therefore results should be interpreted with caution. In this scenario it may be advisable 276
to avoid the use of an antibiotic regimen based solely upon the BHIB enrichment culture if it 277
excludes other possible causative organisms. Further studies are needed to either support or 278
refute these findings. 279
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Conflicts of Interest 363
All authors confirm that there are no conflicts of interest to declare. 364
Funding and Sponsorship 366
No funding or sponsorship was received for this study. 367
... There was also no impact of the type of implicated microorganism, although methicillin-resistant S. aureus and Gram negative bacilli have been reported to be associated with higher rates of failure of arthroscopic lavage [6,7]. All 9 cases in which no culprit microorganism was isolated were in the "success" group, leaving open the legitimate question of possible initial misdiagnosis; interpretation, however, has to be cautious, as their initial clinical presentations were systematically torpid and could correspond to a slow-growth bacterium such as coagulase-negative Staphylococcus or Cutibacterium acnes, which are often difficult to identify, requiring prolonged culture, but which mostly show favorable progression [7,24,25]. ...
Introduction: Referral Centers for Bone and Joint Infection (BJI) were set up to optimize BJI management thanks to multidisciplinary teamwork. The main aim of the present study was to assess the impact of setting up the Western France Bone and Joint Infection Referral Center on arthroscopic treatment of septic arthritis of the shoulder and knee. The secondary aim was to identify other risk factors for failure of this treatment. The null hypothesis was that there was no difference between the "success group" and the "failure group". Material and methods: This single-center retrospective study included 52 patients treated for septic arthritis between January 1, 2000 and December 31, 2013 by arthroscopic joint lavage associated to at least 4 weeks' antibiotic therapy. Exclusion criteria comprised: retrospective diagnosis of rheumatoid arthritis after negative bacteriological analysis, early cessation of antibiotic treatment, and follow-up less than 4 weeks. Failure was defined as non-healing after first-line treatment. The primary endpoint was date of treatment compared to the launch date of the Center in the first quarter of 2010. The influence of pre- and intraoperative criteria related to patient, treatment and microorganism was assessed. Results: At follow-up, 17 patients (32.9%) showed failure of first-line treatment and 5 (9.6%) were non-healed at end of treatment, whatever the re-intervention. The failure rate significantly decreased after setting up the Center, from 42.9% to 11.8% (p=0.03). In the failure group, 70.6% of patients showed immunosuppression, versus 37.2% in the success group (p=0.01). Neither time to surgery (p=1), type of microorganism, or performance of antiseptic lavage (p=0.25) or synovectomy (p=0.62) influenced outcome. Conclusion: Multidisciplinary management of septic arthritis improved treatment success. Level of evidence: III, Retrospective comparative study.
... Moreover, broths may preserve microbial vitality and, when added to samples directly in the operating theater, increase sensitivity from 83 to 95 % (Blackmur et al. 2014). On the other hand, use of broth enrichment may affect specificity by contributing to increase isolate rate of contaminants (Jordan et al. 2015). An alternative strategy may be culture of tissue specimens in blood culture bottles, as described previously for synovial fluid (Peel et al. 2016). ...
Prosthetic joint infection is one of the most severe complication following joint arthroplasty, producing a significant worsening of patient’s quality of life. Management of PJIs requires extended courses of antimicrobial therapy, multiple surgical interventions and prolonged hospital stay, with a consequent economic burden, which is thought to markedly increase in the next years due to the expected burden in total joint arthroplasties. The present review summarizes the present knowledge on microbiological diagnosis of prosthetic joint infections, focusing on aethiological agents and discussing pros and cons of the available strategies for their diagnosis. Intra-operative clinical diagnosis and pathogen identification is considered the diagnostic benchmark, however the presence of bacterial biofilm makes pathogen detection with traditional microbiological techniques highly ineffective. Diagnosis of PJIs is a rather complex challenge for orthopedics and requires a strict collaboration between different specialists: orthopaedics, infectivologists, microbiologists, pathologists and radiologists. Diagnostic criteria have been described by national and international association and scientific societies. Clinicians should be trained on how to use it, but more importantly they should know potential and limitation of the available tests in order to use them appropriately.
Résumé Introduction La création des centres de référence pour les infections ostéoarticulaires (CRIOA) a eu pour but d’optimiser la prise en charge de ce type d’infection, grâce à une concertation multidisciplinaire. L’objectif de cette étude était d’étudier l’influence de la mise en place du Centre de référence des infections ostéoarticulaires du Grand Ouest (CRIOGO) sur le traitement arthroscopique des arthrites septiques (AS) de l’épaule et du genou. L’objectif secondaire était d’identifier d’autres facteurs d’échec de ce traitement. L’hypothèse H0 était l’absence de différence entre le groupe « succès » et le groupe « échec ». Matériel et méthode Cette étude rétrospective monocentrique a inclus 52 patients traités pour une AS entre le 01/01/2000 et le 31/12/2013 par lavage arthroscopique associé à une antibiothérapie d’au moins 4 semaines. Les critères d’exclusion étaient un diagnostic rétrospectif d’arthrite rhumatismale après analyse bactériologique négative, un arrêt précoce du traitement antibiotique et un suivi inférieur à 4 semaines. L’échec était défini comme l’absence de guérison après traitement de première intention. Le critère de jugement principal était la date de la prise en charge par rapport à la date de mise en place du CRIOGO au premier trimestre 2010. L’influence de critères pré- et peropératoires, inhérents au patient, au traitement réalisé et au germe identifié a été recherchée. Résultats À la révision, 17 (32,9 %) patients étaient en échec de traitement de première intention et 5 (9,6 %) n’étaient pas guéris en fin de traitement, quel que soit le type de reprise. Le taux d’échec diminuait significativement après la mise en place du CRIOGO : 42,9 % avant contre 11,8 % après (p = 0,03). Dans le groupe échec, 70,6 % des patients étaient immunodéprimés contre 37,2 % dans le groupe succès (p = 0,01). Ni le délai de prise en charge (p = 1), ni le type de germe, ni un lavage antiseptique (p = 0,25) ou une synovectomie (p = 0,62) n’influençaient le résultat du traitement. Discussion La prise en charge multidisciplinaire des AS permet d’améliorer le succès thérapeutique. Niveau de preuve III, étude rétrospective comparative.
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Prosthetic joint implant surgery (arthroplasty) provides restoration of movement to almost half a million people each year in the USA, with major health and economic benefits. While the majority of recipients experience dramatic relief of preoperative pain at the arthroplasty site, some prostheses fail to achieve this result. This is most commonly the result of aseptic biomechanical failure or prosthetic joint infection (PJI), both of which are associated with significant morbidity. Because the management of PJI differs from that of aseptic failure, it is important to accurately differentiate these two entities. Current laboratory methods for diagnosis of PJI depend on isolation of a pathogen by culture from a clinical specimen (e.g., synovial fluid, periprosthetic tissue). However, as PJI is typically a low organism burden and focal infection caused by commensal micro-organisms, these methods have neither ideal sensitivity nor ideal specificity. Therefore, culture-independent molecular methods have been used to improve the diagnosis of PJI. In the research setting, detection of 16S ribosomal DNA (rDNA) by polymerase chain reaction has been the prime focus of the molecular diagnosis of PJI. In this article, advantages and limitations of conventional and molecular methods for the laboratory diagnosis of PJI are reviewed, as is the use of methods that may improve detection of organisms in the biofilm environment. The diagnosis of PJI remains challenging and a rapid, sensitive and specific method is needed for appropriate surgical and medical treatment decisions in the growing arthroplasty population.
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A prospective study was performed to establish criteria for the microbiological diagnosis of prosthetic joint infection at elective revision arthroplasty. Patients were treated in a multidisciplinary unit dedicated to the management and study of musculoskeletal infection. Standard multiple samples of periprosthetic tissue were obtained at surgery, Gram stained, and cultured by direct and enrichment methods. With reference to histology as the criterion standard, sensitivities, specificities, and likelihood ratios (LRs) were calculated by using different cutoffs for the diagnosis of infection. We performed revisions on 334 patients over a 17-month period, of whom 297 were evaluable. The remaining 37 were excluded because histology results were unavailable or could not be interpreted due to underlying inflammatory joint disease. There were 41 infections, with only 65% of all samples sent from infected patients being culture positive, suggesting low numbers of bacteria in the samples taken. The isolation of an indistinguishable microorganism from three or more independent specimens was highly predictive of infection (sensitivity, 65%; specificity, 99.6%; LR, 168.6), while Gram staining was less useful (sensitivity, 12%; specificity, 98%; LR, 10). A simple mathematical model was developed to predict the performance of the diagnostic test. We recommend that five or six specimens be sent, that the cutoff for a definite diagnosis of infection be three or more operative specimens that yield an indistinguishable organism, and that because of its low level of sensitivity, Gram staining should be abandoned as a diagnostic tool at elective revision arthroplasty.
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Improving diagnosis of prosthetic joint infections (PJIs) has become an increasing challenge due to a steadily rising number of patients with prosthetic implants. Based on a systematic literature search we have ascertained the evidence base for improvement of culture diagnosis. We searched PubMed/MEDLINE using the medical subject heading (MeSH) 'prosthesis-related infections' 1995 through 2010 without further restrictions. An analogous search was conducted for ISI Web of Knowledge. A total of 1409 reports were screened for original results, obtained by methods described in sufficient detail to make replication possible. We gave priority to methods for sample preparation, culture media, culture methods and incubation time. Clinical sensitivity and specificity were calculated where possible. We found evidence to support superiority of cultures obtained from the diluent after sonication of prosthetic implants in comparison with culturing tissue biopsies. Sonication parameters and accessory steps have been studied extensively, and thresholds for significant growth have been defined. Conversely, methods for processing of soft tissue biopsies have been studied to a limited extent. Culture of synovial fluid in blood culture vials has been shown to be more sensitive (90-92%) than intraoperative swab cultures (68-76%) and tissue cultures (77-82%). Formal evaluation of agar media for culturing PJI specimens seemed to be lacking. The polymicrobial nature of PJls supports the routine use of an assortment of media suitable for recovery of fastidious, slow-growing, anaerobic and sublethally damaged bacteria. A number of studies supported an incubation period for up to 14 days. Although we identified evidence-based improvements of culture methods, there is a need for more studies especially with regard to tissue biopsies. Culturing remains an important means to identify and characterize pathogenic micro-organisms and supplements the increasing number of culture-independent assays.
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Clin Microbiol Infect 2011; 17: 1528–1530 The diagnosis of prosthetic joint infection (PJI) in the routine microbiology laboratory is labour-intensive, but semi-automated methods may be appropriate. We prospectively compared four microbiological culture methods on samples taken at prosthetic joint revision surgery. Automated BACTEC blood culture bottles and cooked meat enrichment broth were the most sensitive methods (87% and 83%, respectively, as compared with fastidious anaerobic broth (57%) and direct plates (39%)); all were highly specific (97–100%). To our knowledge, this is the first prospective study aimed at comparing culture methods in routine use in UK clinical laboratories for the diagnosis of PJI.
Periprosthetic joint infection continues to frustrate the medical community. Although the demand for total joint arthroplasty is increasing, the burden of such infections is increasing even more rapidly, and they pose a unique challenge because their accurate diagnosis and eradication can prove elusive. This review describes the current knowledge regarding diagnosis and treatment of periprosthetic joint infection. A number of tools are available to aid in establishing a diagnosis of periprosthetic joint infection. These include the erythrocyte sedimentation rate, serum C-reactive protein concentration, synovial white blood-cell count and differential, imaging studies, tissue specimen culturing, and histological analysis. Multiple definitions of periprosthetic joint infection have been proposed but there is no consensus. Tools under investigation to diagnose such infections include the C-reactive protein concentration in the joint fluid, point-of-care strip tests for the leukocyte esterase concentration in the joint fluid, and other molecular markers of periprosthetic joint infection. Treatment options include irrigation and debridement with prosthesis retention, one-stage prosthesis exchange, two-stage prosthesis exchange with intervening placement of an antibiotic-loaded spacer, and salvage treatments such as joint arthrodesis and amputation. Treatment selection is dependent on multiple factors including the timing of the symptom onset, patient health, the infecting organism, and a history of infection in the joint. Although prosthesis retention has the theoretical advantages of decreased morbidity and improved return to function, two-stage exchange provides a lower rate of recurrent infection. As the burden of periprosthetic joint infection increases, the orthopaedic and medical community should become more familiar with the disease. It is hoped that the tools currently under investigation will aid clinicians in diagnosing periprosthetic joint infection in an accurate and timely fashion to allow appropriate treatment. Given the current knowledge and planned future research, the medical community should be prepared to effectively manage this increasingly prevalent disease.
Dislocation and infection are common complications of total hip arthroplasty (THA). This study evaluated the correlation between the number of revision THAs and the incidence of these complications. Data were obtained from 749 revision THAs. Average follow-up was 13.2 ± 5.9 years. Patients were grouped as first, second, third, and fourth or greater revision THA. Dislocation rates (5.68%, 7.69%, 8.33%, and 27.45%) and infection rates (1.35%, 1.92%, 2.5%, and 7.84%) in the first, second, third, and fourth or greater groups, respectively, correlated directly with the revision number and were highest (P < .001) in the fourth or greater group. Dislocation and infection are exponentially correlated with the number of revision THA. From the fourth revision onward, those risks are multiplied.
We evaluated the performance of the BACTEC Peds Plus bottles for the detection of bacteria in 186 tissue samples obtained from orthopedic infections. BACTEC Peds Plus bottles led to bacterial detection in 69% of these samples against less than 53% for each of the other types of conventional media (P < 0.05). For some patients, the time of detection of pathogens was lower with the BACTEC Peds Plus bottles than with the conventional media.