Université d'Aix-Marseille, Unité des rickettsies, URMITE CNRS-IRD, Faculté de médecine, Marseille, France.
Clin Microbiol Infect. 2012 Dec;18(12):1168-75. doi: 10.1111/1469-0691.12020.
Microbiological culture is the conventional method for establishing the diagnosis in implant-associated bone and joint infection, but it may lack both specificity and sensitivity. Molecular diagnosis has been an important step in the diagnosis of infectious diseases. We review the principles and the role of molecular diagnosis in improving the aetiological diagnosis of implant-associated bone and joint infection. Currently, molecular diagnosis mainly includes conventional broad-range PCR and specific PCR assays. These tools are efficient, but several pitfalls exist that necessitate rigour in all steps of the process. In implant-associated bone and joint infection, molecular assays have been shown to be useful in complementing culture techniques to identify microorganisms when patients have previously received antibiotics or in the presence of fastidious microorganisms. Broad-range PCR targeting the 16S rRNA sequence followed by sequencing must be performed in culture-negative specimens when infection is suspected on the basis of clinical signs and symptoms or inflammatory syndrome. This molecular tool has allowed not only increasing identification of anaerobic bacteria, such as Finegoldia magna, but also the discovery of the role of Tropheryma whipplei, an aetiological agent of implant-associated bone and joint infection in patients without Whipple's disease. Real-time pathogen-specific PCR assays performed in a closed system are more sensitive and specific than broad-range PCR, but each assay is typically able to detect only a single microorganism. These assays should be performed to confirm the identification provided by broad-spectrum PCR, and also when broad-range PCR fails to detect a microorganism despite efficient DNA extraction.
微生物培养是诊断植入物相关骨和关节感染的常规方法,但它可能缺乏特异性和敏感性。分子诊断一直是传染病诊断的重要步骤。我们回顾了分子诊断在提高植入物相关骨和关节感染病因诊断中的原理和作用。目前,分子诊断主要包括常规广谱 PCR 和特定 PCR 检测。这些工具效率很高,但存在一些需要在整个过程中严格控制的陷阱。在植入物相关骨和关节感染中,分子检测已被证明在以下情况下有助于补充培养技术,以识别接受过抗生素治疗的患者或存在难培养微生物时的微生物:基于临床症状和炎症综合征怀疑感染,但培养结果为阴性。当怀疑感染时,必须对培养阴性标本进行靶向 16S rRNA 序列的广谱 PCR 检测,然后进行测序。这种分子工具不仅提高了对厌氧菌(如金氏金杆菌)的鉴定能力,而且还发现了特罗非马属(Tropheryma whipplei)在没有惠特利病的植入物相关骨和关节感染患者中的作用。在封闭系统中进行的实时病原体特异性 PCR 检测比广谱 PCR 更敏感和特异,但每种检测通常只能检测一种微生物。这些检测应在以下情况下进行:确认广谱 PCR 提供的鉴定结果;以及尽管进行了有效的 DNA 提取,但广谱 PCR 仍未能检测到微生物时。