Dupieux Céline, Descours Ghislaine, Verhoeven Paul, Grattard Florence, Benito Yvonne, Vandenesch François, Cazorla Céline, Ferry Tristan, Lustig Sébastien, Boyer Bertrand, Boisset Sandrine, Carricajo Anne, Laurent Frédéric, Pirla Investigator Group /
Laboratoire de Bactériologie, Institut des Agents Infectieux, Hôpital de la Croix-Rousse, Hospices Civils de Lyon, Lyon, 69004, France.
Laboratoire de Bactériologie, CHU de Saint-Étienne, Saint-Étienne, 42055, France.
J Bone Jt Infect. 2024 Feb 6;9(1):37-48. doi: 10.5194/jbji-9-37-2024. eCollection 2024.
No consensus exists about the techniques to use for microbiological diagnosis of bone and joint infections (BJIs). The objective herein was to define an algorithm to optimize BJI diagnosis in adults using various bacteriological methods on synovial fluid samples. This prospective multi-center study included 423 synovial fluids collected from adult patients with suspected BJIs. Culture (using five solid media, an enrichment broth, and blood culture bottles), universal 16S rRNA PCR followed by Sanger sequencing, and seven specific bacterial PCRs were systematically performed. Combinations of methods were compared to arrive at the optimized algorithm. Among 423 synovial fluids, 242 infections were diagnosed (57.2 %): 213 mono- and 29 poly-microbial for a total of 284 bacteria (staphylococci at 54.6 %, streptococci-enterococci at 16.5 %, Gram-negative bacilli at 15.5 %, anaerobic species at 8.8 %). Comparing culture techniques, blood culture bottles had the highest sensitivity (67.6 % for pediatric and 63.9 % for anaerobic bottles) but are not sufficient alone and require being combined with solid media. The 16S rDNA PCR detected only 52.3 % of the bacteria, whereas specific PCRs had a higher sensitivity ( spp. at 66.2 %, at 85.2 %, spp. at 91.2 %). Based on these results, an algorithm was proposed associating three solid media; inoculation into blood culture bottles; and 16S, spp., and spp. PCRs, which would have detected 90.5 % of bacteria in the present cohort versus 79.2 % using all culture techniques on synovial fluid. This prospective study shows that a combination of culture and molecular methods on synovial fluids allows the optimization of bacterial detection.
对于骨与关节感染(BJI)的微生物学诊断应采用何种技术,目前尚无共识。本文的目的是制定一种算法,以利用滑膜液样本上的各种细菌学方法优化成人BJI的诊断。这项前瞻性多中心研究纳入了423份从疑似BJI的成年患者采集的滑膜液。系统地进行了培养(使用五种固体培养基、一种增菌肉汤和血培养瓶)、通用16S rRNA PCR随后进行桑格测序,以及七种特定细菌PCR检测。对各种方法的组合进行比较,以得出优化算法。在423份滑膜液中,诊断出242例感染(57.2%):213例为单一微生物感染,29例为多微生物感染,共检出284种细菌(葡萄球菌占54.6%,链球菌 - 肠球菌占16.5%,革兰氏阴性杆菌占15.5%,厌氧菌占8.8%)。比较培养技术发现,血培养瓶的敏感性最高(儿科血培养瓶为67.6%,厌氧菌血培养瓶为63.9%),但单独使用并不充分,需要与固体培养基联合使用。16S rDNA PCR仅检测到52.3%的细菌,而特定PCR的敏感性更高([具体菌种1]为66.2%,[具体菌种2]为85.2%,[具体菌种3]为91.2%)。基于这些结果,提出了一种算法组合,包括三种固体培养基;接种到血培养瓶中;以及16S、[具体菌种1]和[具体菌种2]的PCR检测,该算法在本队列中可检测到90.5%的细菌,而仅对滑膜液使用所有培养技术的检测率为79.2%。这项前瞻性研究表明,滑膜液培养和分子方法的结合可优化细菌检测。