Detection of bacteria with molecular methods in prosthetic joint infection: sonication fluid better than periprosthetic tissue

Rak, Mitja; Kavčič, Martina; Trebše, Rihard; Cör, Andrej

doi:10.3109/17453674.2016.1165558

Cited by 50 publications

(24 citation statements)

References 27 publications

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“…Sonication of prostheses has proved to be reliable, effective, and reproducible [23,31]. Sonicates of implants have been used in both conventional and molecular microbiological methods, and more frequently than periprosthetic tissues as PCR material for PJI diagnosis, with superior sensitivity [27,32,33]. Our results agree with these studies as the sonicate PCR sensitivity was 64.3% and 71.4% for the conventional and viability PCR respectively compared to 15.4% and 23% for the periprosthetic tissues.…”

Section: Discussionsupporting

confidence: 84%

Propidium monoazide–polymerase chain reaction for detection of residual periprosthetic joint infection in two-stage revision

et al. 2019

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False negative culture results in periprosthetic joint infection (PJI) are not uncommon particularly when patients have received long term antibiotics. Polymerase chain reaction (PCR) has a lower specificity partly due to detection of residual DNA from dead bacteria. Propidium monoazide (PMA) prevents DNA from dead bacteria from being amplified during the PCR. This study aimed to determine the role of PMA in PCR for diagnosis of PJI. Clinical samples were tested by PCR with and without prior treatment with PMA and compared to conventional microbiological culture. The PCR assay included genus-specific primers for staphylococci and enterococci and species-specific primers for Cutibacterium acnes. The validated conditions of PMA treatment used in this study were 20 μM concentration and 5 and 10 min of dark incubation and photo-activation respectively. 202 periprosthetic tissues and explanted prostheses from 60 episodes in 58 patients undergoing revision arthroplasties for either PJI or non-infective causes were tested, by culture, PCR, and PMA-PCR. 14 of the 60 episodes satisfied the Musculoskeletal Infection Society (MSIS) criteria for PJI and 46 did not. Sensitivity of culture, PCR, and PMA-PCR were 50%, 71%, and 79% respectively. Specificities were 98%, 72%, and 89% respectively. All figures were calculated for episodes rather than samples. PMA-PCR enhanced both the specificity and the sensitivity of PCR. It has the potential to detect residual bacterial viability prior to reimplantation in the two-stage revision for PJI.

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Section: Discussionsupporting

confidence: 84%

Propidium monoazide–polymerase chain reaction for detection of residual periprosthetic joint infection in two-stage revision

et al. 2019

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“…A study that compared cultures obtained from sonication fluid and PCR for PJI determined that in patients with prior antibiotic therapy mPCR was positive in 100%, whereas cultures from sonication only in 42% 32 . In further studies could be shown another advantage that PCR of prosthesis sonication samples is more sensitive than tissue culture for the microbiologic diagnosis of PJI and provides same-day diagnosis with definition of microbiology 34 , 35 . In a recent study was stated, that PCR and microarray-based platform provide the attractive possibility of faster bacterial diagnosis than with routine culture, and the molecular methods were most helpful in PJI diagnostics during ongoing antimicrobial treatment 36 .…”

Section: Discussionmentioning

confidence: 99%

Are There Benefits In Early Diagnosis Of Prosthetic Joint Infection With Multiplex Polymerase Chain Reaction?

Lausmann

Zahár

Citak

et al. 2017

J. Bone Joint Infect.

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Purpose Identification of bacteria and susceptibility are fundamental in periprosthetic joint infection (PJI). Especially in the case of systemic inflammatory response syndrome (SIRS) rapid detection of pathogens is essential for proper therapy. Bacterial cultures are time consuming. The polymerase chain reaction (PCR) is a non-culture molecular method and is able to rapidly identify pathogens and their resistance genes. Multiplex PCR (mPCR) can amplify several different DNA sequences simultaneously. The aim of this study was to show the value of mPCR for early diagnosis of PJI.Methods 60 patients undergoing total hip or knee revisions were recruited in this prospective single-centre-study. Three groups were created: 26 patients with aseptic loosening (negative control), 26 patients with chronic PJI, and 8 patients with acute PJI/SIRS. We compared the results of joint aspirates obtained intraoperatively investigated by mPCR with the microbiology results of tissue specimens.Results The overall sensitivity of mPCR was 78.8% (95% CI, 61.1 - 91.0%), the specificity was 100% (95% CI, 87.2 - 100%), the negative predictive value was 79.4% (95% CI, 62.1 - 91.3%), the positive predictive value was 100% (95% CI, 86.8 - 100%), and the overall accuracy was 88.3% (95% CI, 77.4 - 95.2%). The overall accuracy in acute infections/SIRS (87.5%) was greater than in late chronic PJI (76.9%). In PJI the mPCR was able to provide the results within 5 hours whereas the mean time for cultures was 6.4 days.Conclusions Multiplex PCR is a reliable diagnostic tool in PJI management, especially in acute cases complicated with SIRS. Early diagnosis within several hours is possible, targeted antibiotic treatment can be started promptly.

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“…The great benefit of this PCR technique in diagnosing PJI was documented especially in patients who received antibiotic therapy close to surgery or for a longer period 7 8 . Studies suggest that NAT of sonication fluid may further increase sensitivity and specificity 37 . Unfortunately, this technique is not routinely available in laboratories, due to its time-consuming workflow.…”

Section: Discussionmentioning

confidence: 99%

“…There are certain limitations to the PCR technique in general: NAT detects DNA with no differentiation between viable and non-viable bacteria, making interpretation of the results difficult. Broad-range PCR will only detect ribosomal 16S ribonucleic acid (16S rRNA), not differentiating between pathogens 37 . More specific systems do not routinely detect gene-encoded antibiotic resistance markers.…”

Section: Discussionmentioning

confidence: 99%

Novel Diagnostics in Revision Arthroplasty: Implant Sonication and Multiplex Polymerase Chain Reaction

Hischebeth

Gravius

Buhr

et al. 2017

JoVE

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In orthopedic patients, foreign body-associated infections, especially periprosthetic joint infections (PJIs), are a devastating complication of arthroplasty. Infection requires complex treatment, may result in long hospitalization and causes considerable costs. Multiple surgical revisions can be necessary in these patients, with a loss in function as well as in quality of life.The routine preoperative diagnostics include blood examination for C-reactive protein (CRP) and other biomarkers, as well as joint aspirate analysis for cell count, differentiation, and culture. Intraoperative specimens for histology and microbiology are also standard procedure. The microbiological examination of removed implants with sonication, in combination with the implementation of molecular biology techniques in microbiology, represent two novel techniques currently employed to enhance the differential diagnostics of PJI.We present here the step-wise procedure of analyzing joint aspirate and sonication fluid, using a cartridge-based multiplex polymerase chain reaction (PCR) system. Results were matched against conventional cultures and consensus criteria for PJI. Conventional microbiological cultures from tissue biopsies, joint aspirate and sonication fluid showed a sensitivity of 66.7%, 66.7%, and 88.9%, respectively, and a specificity of 82.3%, 54.6%, and 61.5%, respectively. The PCR diagnostic of the sonication fluid and the joint fluid showed a sensitivity of 50.0% and 55.6%, respectively, and both a specificity of 100.0%. Both PCR diagnostics combined had a sensitivity of 66.7% and a specificity of 100.0%. The multiplex PCR therefore presents a rapid diagnostic tool with moderate sensitivity but high specificity in diagnosing PJI.

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Detection of bacteria with molecular methods in prosthetic joint infection: sonication fluid better than periprosthetic tissue

Cited by 50 publications

References 27 publications

Propidium monoazide–polymerase chain reaction for detection of residual periprosthetic joint infection in two-stage revision

Propidium monoazide–polymerase chain reaction for detection of residual periprosthetic joint infection in two-stage revision

Are There Benefits In Early Diagnosis Of Prosthetic Joint Infection With Multiplex Polymerase Chain Reaction?

Novel Diagnostics in Revision Arthroplasty: Implant Sonication and Multiplex Polymerase Chain Reaction

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