Identification of the same species in at least two intra-operative samples for prosthetic joint infection diagnostics yields the best results with broad-range polymerase chain reaction

Rak, Mitja; Barlič–Maganja, Darja; Kavčič, Martina; Trebše, Rihard; Cör, Andrej

doi:10.1007/s00264-014-2552-2

Cited by 11 publications

(8 citation statements)

References 23 publications

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“…In 6 of 9 patients with AF where only periprosthetic tissue was BR-PCR-positive (patients 10, 14, 31, 36, 39, and 79), the bacteria identified were considered insignificant based on criteria for interpretation of BR-PCR results of periprosthetic tissue analysis (Rak et al. 2015 ). The bacteria identified in periprosthetic tissue of the remaining 3 patients (patients 16, 17, and 63) were considered significant based on the criteria for interpretation of BR-PCR results.…”

Section: Resultsmentioning

confidence: 99%

“…Only 3 of 9 bacteria were considered to be significant based on criteria for interpretation of BR-PCR results of periprosthetic tissue analysis (Rak et al. 2015 ). However, the origin of these bacteria is not known for 2 of the 3 cases (patients 16 and 17), because culture of periprosthetic tissue was negative.…”

Section: Discussionmentioning

confidence: 99%

“…Aliquots of 1 mL were prepared for molecular methods, but the rest of the sample was used for routine culture (Rak et al. 2013 , 2015 ).…”

Section: Methodsmentioning

confidence: 99%

“…Molecular analyses of periprosthetic tissue samples were done as previously described (Rak et al. 2013 , 2015 ). Briefly, total DNA was extracted with a PureLink Genomic DNA Mini Kit (Invitrogen) according to the manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2016

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Background and purposeThe correct diagnosis of prosthetic joint infection (PJI) can be difficult because bacteria form a biofilm on the surface of the implant. The sensitivity of culture from sonication fluid is better than that from periprosthetic tissue, but no comparison studies using molecular methods on a large scale have been performed. We assessed whether periprosthetic tissue or sonication fluid should be used for molecular analysis.Patients and methodsImplant and tissue samples were retrieved from 87 patients who underwent revision operation of total knee or total hip arthroplasty. Both sample types were analyzed using broad-range (BR-) PCR targeting the 16S rRNA gene. The results were evaluated based on the definition of periprosthetic joint infection from the Workgroup of the Musculoskeletal Infection Society.ResultsPJI was diagnosed in 29 patients, whereas aseptic failure was diagnosed in 58 patients. Analysis of sonication fluid using BR-PCR detected bacteria in 27 patients, whereas analysis of periprosthetic tissue by BR-PCR detected bacteria in 22 patients. In 6 of 7 patients in whom BR-PCR analysis of periprosthetic tissue was negative, low-virulence bacteria were present. The sensitivity and specificity values for periprosthetic tissue were 76% and 93%, respectively, and the sensitivity and specificity values for sonication fluid were 95% and 97%.InterpretationOur results suggest that sonication fluid may be a more appropriate sample than periprosthetic tissue for BR-PCR analysis in patients with PJI. However, further investigation is required to improve detection of bacteria in patients with so-called aseptic failure.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…Aliquots of 1 mL were prepared for molecular methods, but the rest of the sample was used for routine culture (Rak et al. 2013 , 2015 ).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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

et al. 2016

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“…The PCR assay included genus-specific primers for staphylococci [19] and enterococci [20] and species-specific primers for Cutibacterium acnes [21]. The human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) housekeeping gene [22] was amplified as an internal control of DNA extraction and possible PCR inhibition (Table 1). Separate PCR assays were used for each primer pair.…”

Section: Application Of Optimised Pma In Clinical Samplesmentioning

confidence: 99%

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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Microbiological Diagnosis of Implant-Related Infections: Scientific Evidence and Cost/Benefit Analysis of Routine Antibiofilm Processing

Drago

Vecchi

2016

Advances in Experimental Medicine and Biology

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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.

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Identification of the same species in at least two intra-operative samples for prosthetic joint infection diagnostics yields the best results with broad-range polymerase chain reaction

Abstract: For diagnostics of PJI with BR-PCR the criterion of the same bacterial species in at least two specimens of periprosthetic tissue from the same patient should be used for interpretation of positive results.

Cited by 11 publications

References 23 publications

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

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

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

Microbiological Diagnosis of Implant-Related Infections: Scientific Evidence and Cost/Benefit Analysis of Routine Antibiofilm Processing

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