Comparison of culture and broad-range polymerase chain reaction methods for diagnosing periprosthetic joint infection: analysis of joint fluid, periprosthetic tissue, and sonicated fluid

Objectives The objective of this study was to develop a test for the rapid (within 25 minutes) intraoperative detection of bacteria from synovial fluid to diagnose periprosthetic joint infection (PJI). Methods The 16s rDNA test combines a polymerase chain reaction (PCR) for amplification of 16s rDNA with a lateral flow immunoassay in one fully automated system. The synovial fluid of 77 patients undergoing joint aspiration or primary or revision total hip or knee surgery was prospectively collected. The cohort was divided into a proof-of-principle cohort (n = 17) and a validation cohort (n = 60). Using the proof-of-principle cohort, an optimal cut-off for the discrimination between PJI and non-PJI samples was determined. PJI was defined as detection of the same bacterial species in a minimum of two microbiological samples, positive histology, and presence of a sinus tract or intra-articular pus. Results The 16s rDNA test proved to be very robust and was able to provide a result in 97% of all samples within 25 minutes. The 16s rDNA test was able to diagnose PJI with a sensitivity of 87.5% and 82%, and a specificity of 100% and 89%, in the proof-of-principle and validation cohorts, respectively. The microbiological culture of synovial fluid achieved a sensitivity of 80% and a specificity of 93% in the validation cohort. Conclusion The 16s rDNA test offers reliable intraoperative detection of all bacterial species within 25 minutes with a sensitivity and specificity comparable with those of conventional microbiological culture of synovial fluid for the detection of PJI. The 16s rDNA test performance is independent of possible blood contamination, culture time and bacterial species. Cite this article: V. Janz, J. Schoon, C. Morgenstern, B. Preininger, S. Reinke, G. Duda, A. Breitbach, C. F. Perka, S. Geissler. Rapid detection of periprosthetic joint infection using a combination of 16s rDNA polymerase chain reaction and lateral flow immunoassay: A Pilot Study. Bone Joint Res 2018;7:12–19. DOI: 10.1302/2046-3758.71.BJR-2017-0103.R2.

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Detecting the Presence of Bacterial DNA and RNA by Polymerase Chain Reaction to Diagnose Suspected Periprosthetic Joint Infection after Antibiotic Therapy

Orthopaedic Surgery ◽

10.1111/os.12359 ◽

2018 ◽

Vol 10 (1) ◽

pp. 40-46 ◽

Cited By ~ 4

Author(s):

Xin-yu Fang ◽

Wen-bo Li ◽

Chao-fan Zhang ◽

Zi-da Huang ◽

Hui-yi Zeng ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Antibiotic Therapy ◽

Periprosthetic Joint Infection ◽

Joint Infection ◽

Chain Reaction ◽

Bacterial Dna ◽

Dna And Rna ◽

Polymerase Chain

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A new multiplex real-time polymerase chain reaction assay for the diagnosis of periprosthetic joint infection

Modern Rheumatology ◽

10.1080/14397595.2017.1295825 ◽

2017 ◽

Vol 27 (6) ◽

pp. 1072-1078 ◽

Cited By ~ 6

Author(s):

Masaki Kawamura ◽

Naomi Kobayashi ◽

Yutaka Inaba ◽

Hyonmin Choe ◽

Taro Tezuka ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Periprosthetic Joint Infection ◽

Polymerase Chain Reaction Assay ◽

Joint Infection ◽

Chain Reaction ◽

Polymerase Chain

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Diagnosis of Periprosthetic Joint Infection Using Polymerase Chain Reaction: An Updated Systematic Review and Meta-Analysis

Surgical Infections ◽

10.1089/sur.2018.014 ◽

2018 ◽

Vol 19 (6) ◽

pp. 555-565 ◽

Cited By ~ 10

Author(s):

Yang Jun ◽

Liu Jianghua

Keyword(s):

Systematic Review ◽

Polymerase Chain Reaction ◽

Periprosthetic Joint Infection ◽

Meta Analysis ◽

Joint Infection ◽

Chain Reaction ◽

Polymerase Chain

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

Molecular Biology Reports ◽

10.1007/s11033-019-05092-z ◽

2019 ◽

Vol 46 (6) ◽

pp. 6463-6470

Author(s):

Mohamed Askar ◽

Mariam Sajid ◽

Yassar Nassif ◽

Waheed Ashraf ◽

Brigitte Scammell ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Periprosthetic Joint Infection ◽

Joint Infection ◽

Propidium Monoazide ◽

Chain Reaction ◽

Two Stage ◽

Specific Primers ◽

Musculoskeletal Infection ◽

Cutibacterium Acnes ◽

Polymerase Chain

Abstract 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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