Aims The aim of this study was to evaluate the performance of metagenomic next-generation sequencing (mNGS) in detecting pathogens from synovial fluid of prosthetic joint infection (PJI) patients. Methods A group of 75 patients who underwent revision knee or hip arthroplasties were enrolled prospectively. Ten patients with primary arthroplasties were included as negative controls. Synovial fluid was collected for mNGS analysis. Optimal thresholds were determined to distinguish pathogens from background microbes. Synovial fluid, tissue, and sonicate fluid were obtained for culture. Results A total of 49 PJI and 21 noninfection patients were finally included. Of the 39 culture-positive PJI cases, mNGS results were positive in 37 patients (94.9%), and were consistent with culture results at the genus level in 32 patients (86.5%) and at the species level in 27 patients (73.0%). Metagenomic next-generation sequencing additionally identified 15 pathogens from five culture-positive and all ten culture-negative PJI cases, and even one pathogen from one noninfection patient, while yielding no positive findings in any primary arthroplasty. However, seven pathogens identified by culture were missed by mNGS. The sensitivity of mNGS for diagnosing PJI was 95.9%, which was significantly higher than that of comprehensive culture (79.6%; p = 0.014). The specificity is similar between mNGS and comprehensive culture (95.2% and 95.2%, respectively; p = 1.0). Conclusion Metagenomic next-generation sequencing can effectively identify pathogens from synovial fluid of PJI patients, and demonstrates high accuracy in diagnosing PJI. Cite this article: Bone Joint Res 2020;9(7):440–449.

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The purpose of this study was to explore the role of mNGS in the diagnosis of PJI by detecting the presence of bacteria in periprosthetic tissues. Methods: Patients who were suspected of PJI and underwent surgery in our center were enrolled. Microbial culture and mNGS were performed on periprosthetic tissues collected intraoperatively. Patient age, body mass index (BMI), laboratory tests, microbial culture and mNGS results were recorded. The diagnostic performance of microbial culture and mNGS in the diagnosis of PJI was compared. Results: A total of 44 patients were included. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of periprosthetic tissue microbial culture in the diagnosis of PJI were 72.72%, 77.27%, 76.19%, 73.91% and 75%, respectively. The sensitivity, specificity, PPV, NPV and accuracy of periprosthetic tissue mNGS in the diagnosis of PJI were 95.45%, 90.91%, 91.3%, 95.24% and 93.18%, respectively. The sensitivity and specificity of mNGS in the diagnosis of PJI were superior to those of microbial culture. Conclusion: Periprosthetic tissue can be used as an alternative to synovial fluid and sonication fluid, as mNGS can improve the diagnosis of PJI. mNGS of the periprosthetic tissues showed higher sensitivity than microbial culture.

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Preoperative diagnosis is important for patients who need revision surgery due to PJI. Microbial culture plays an important role in PJI diagnosis, but the sensitivity of cultures is low when the sample amount is limited or when a patient is treated with antibiotics before sample collection. In this study, metagenomic next-generation sequencing (mNGS) was used to detect bacteria in preoperative puncture synovial fluid samples from patients with suspected PJI, and the preoperative and intraoperative culture results were compared to estimate its diagnostic efficiency. Methods: From July 2016 to December 2018, patients with suspected PJI who underwent prosthetic joint revision surgery were included, and the results of those who had been tested by preoperative synovial fluid culture and mNGS were obtained. The demographic characteristics, medical history, laboratory test results, culture results, and mNGS results of each patient were recorded. Then, the efficiency of preoperative synovial fluid mNGS was compared to that of synovial fluid culture for diagnosing PJI. Results: A total of 37 patients were included, and 24 patients (25 joints) were diagnosed with PJI. The sensitivity, specificity, and accuracy of preoperative synovial fluid mNGS were 92%, 91.7%, and 83.7%, respectively. The sensitivity, specificity, and accuracy of preoperative synovial fluid culture were 52%, 91.7%, and 43.7%, respectively. Conclusions: With a low volume of synovia (1 ml), mNGS can be performed with higher sensitivity and specificity compared to synovial culture. Thus, mNGS can be a useful supplemental method to improve diagnostic efficiency during the preoperative period.