Department of Medical Microbiology and Infection Prevention, University of Groningen, University Medical Center Groningen (UMCG), Hanzeplein 1, 9700RB, Groningen, The Netherlands.
Department of Orthopaedics, University of Groningen, University Medical Center Groningen (UMCG), Groningen, The Netherlands.
Eur J Nucl Med Mol Imaging. 2024 Aug;51(10):2988-2997. doi: 10.1007/s00259-023-06499-4. Epub 2023 Nov 14.
Staphylococcus aureus is the most common and impactful multi-drug resistant pathogen implicated in (periprosthetic) joint infections (PJI) and fracture-related infections (FRI). Therefore, the present proof-of-principle study was aimed at the rapid detection of S. aureus in synovial fluids and biofilms on extracted osteosynthesis materials through bacteria-targeted fluorescence imaging with the 'smart-activatable' DNA-based AttoPolyT probe. This fluorogenic oligonucleotide probe yields large fluorescence increases upon cleavage by micrococcal nuclease, an enzyme secreted by S. aureus.
Synovial fluids from patients with suspected PJI and extracted osteosynthesis materials from trauma patients with suspected FRI were inspected for S. aureus nuclease activity with the AttoPolyT probe. Biofilms on osteosynthesis materials were imaged with the AttoPolyT probe and a vancomycin-IRDye800CW conjugate (vanco-800CW) specific for Gram-positive bacteria.
38 synovial fluid samples were collected and analyzed. Significantly higher fluorescence levels were measured for S. aureus-positive samples compared to, respectively, other Gram-positive bacterial pathogens (p < 0.0001), Gram-negative bacterial pathogens (p = 0.0038) and non-infected samples (p = 0.0030), allowing a diagnosis of S. aureus-associated PJI within 2 h. Importantly, S. aureus-associated biofilms on extracted osteosynthesis materials from patients with FRI were accurately imaged with the AttoPolyT probe, allowing their correct distinction from biofilms formed by other Gram-positive bacteria detected with vanco-800CW within 15 min.
The present study highlights the potential clinical value of the AttoPolyT probe for fast and accurate detection of S. aureus infection in synovial fluids and biofilms on extracted osteosynthesis materials.
金黄色葡萄球菌是引起（假体周围）关节感染（PJI）和骨折相关感染（FRI）的最常见且影响最大的多药耐药病原体。因此，本原理验证研究旨在通过使用针对细菌的荧光成像技术，即“智能激活”基于 DNA 的 AttoPolyT 探针，快速检测滑液和提取的内固定材料上的生物膜中的金黄色葡萄球菌。这种荧光寡核苷酸探针在被金黄色葡萄球菌分泌的微球菌核酸酶切割时会产生较大的荧光增强。
使用 AttoPolyT 探针检测疑似 PJI 患者的滑液和疑似 FRI 患者的提取内固定材料中的金黄色葡萄球菌核酸酶活性。使用 AttoPolyT 探针和针对革兰氏阳性菌的万古霉素-IRDye800CW 缀合物（vanco-800CW）对骨固定材料上的生物膜进行成像。
共收集和分析了 38 份滑液样本。与其他革兰氏阳性细菌病原体（p<0.0001）、革兰氏阴性细菌病原体（p=0.0038）和非感染样本（p=0.0030）相比，金黄色葡萄球菌阳性样本的荧光水平显著升高，这使得在 2 小时内可以诊断出金黄色葡萄球菌相关性 PJI。重要的是，使用 AttoPolyT 探针可以准确地对 FRI 患者提取的内固定材料上的金黄色葡萄球菌相关生物膜进行成像，在 15 分钟内可以将其与用 vanco-800CW 检测到的其他革兰氏阳性菌形成的生物膜区分开来。
本研究强调了 AttoPolyT 探针在快速准确检测滑液和提取的内固定材料上的金黄色葡萄球菌感染方面的潜在临床价值。
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