Portillo Maria Eugenia, Sancho Ignacio
Department of Clinical Microbiology, University Hospital of Navarra, Institute of Healthcare Research of Navarra IdiSNA, 31008 Pamplona, Spain.
Department of Orthopedics and Trauma Surgery, University Hospital of Navarra, 31008 Pamplona, Spain.
Diagnostics (Basel). 2023 Feb 20;13(4):809. doi: 10.3390/diagnostics13040809.
A significant number of prosthetic joint infections (PJI) are culture-negative and/or misinterpreted as aseptic failures in spite of the correct implementation of diagnostic culture techniques, such as tissue sample processing in a bead mill, prolonged incubation time, or sonication of removed implants. Misinterpretation may lead to unnecessary surgery and needless antimicrobial treatment. The diagnostic value of non-culture techniques has been investigated in synovial fluid, periprosthetic tissues, and sonication fluid. Different feasible improvements, such as real-time technology, automated systems and commercial kits are now available to support microbiologists. In this review, we describe non-culture techniques based on nucleic acid amplification and sequencing methods. Polymerase chain reaction (PCR) is a frequently used technique in most microbiology laboratories which allows the detection of a nucleic acid fragment by sequence amplification. Different PCR types can be used to diagnose PJI, each one requiring the selection of appropriate primers. Henceforward, thanks to the reduced cost of sequencing and the availability of next-generation sequencing (NGS), it will be possible to identify the whole pathogen genome sequence and, additionally, to detect all the pathogen sequences present in the joint. Although these new techniques have proved helpful, strict conditions need to be observed in order to detect fastidious microorganisms and rule out contaminants. Specialized microbiologists should assist clinicians in interpreting the result of the analyses at interdisciplinary meetings. New technologies will gradually be made available to improve the etiologic diagnoses of PJI, which will remain an important cornerstone of treatment. Strong collaboration among all specialists involved is essential for the correct diagnosis of PJI.
尽管正确实施了诊断培养技术,如在珠磨仪中处理组织样本、延长培养时间或对取出的植入物进行超声处理,但仍有相当数量的人工关节感染(PJI)培养结果为阴性和/或被误诊为无菌性失败。误诊可能导致不必要的手术和不必要的抗菌治疗。非培养技术在滑液、假体周围组织和超声处理液中的诊断价值已得到研究。现在有不同的可行改进方法,如实时技术、自动化系统和商业试剂盒来支持微生物学家。在本综述中,我们描述基于核酸扩增和测序方法的非培养技术。聚合酶链反应(PCR)是大多数微生物实验室常用的技术,它通过序列扩增来检测核酸片段。不同类型的PCR可用于诊断PJI,每种类型都需要选择合适的引物。今后,由于测序成本的降低和新一代测序(NGS)的可用性,将有可能鉴定整个病原体基因组序列,此外,还能检测关节中存在的所有病原体序列。尽管这些新技术已被证明是有帮助的,但为了检测苛养微生物并排除污染物,仍需遵守严格的条件。专业微生物学家应在跨学科会议上协助临床医生解释分析结果。新技术将逐渐应用于改善PJI的病因诊断,病因诊断仍将是治疗的重要基石。所有相关专家之间的密切合作对于PJI的正确诊断至关重要。