关节假体术后感染的早期诊断和基因分型研究

The early diagnosis of prosthetic joint infection (PJI) was just the clinical puzzles that hasn't solved yet. The pathogenesis of PJI is typically caused by microorganisms growing in structures, known as biofilms and microorganisms are typically present in a biofilm on the surface of the prosthesis. The management of PJIs remains a clinical challenge particularly when standard aerobic and anaerobic culture techniques fail to isolate the causative pathogen, so called ‘culture negative prosthetic joint infection’. Culturing of clinical samples is the standard method used for the microbiologic diagnosis of PJI, but this method is neither sensitive nor specific. With the development of pathogen epidemiology and genetic research theory of PJI, we hypothesized that detecting general and specific genotype would improve the microbiologic diagnosis of PJI at early stage. 16S rRNA was selected as a general gene in early diagnosis of PJI in order to improve the diagnostic sensitivity, and then the specific genotype (ICA D, Eno, SAR A, agr, Mec A and Mre B gene) were combined detected to improve the diagnostic specificity. It was verified through in vitro and animal experiment. If our new strategy of the diagnosis is a sensitive and specific test for the diagnosis of PJI in patients at early stage, it will provide experimental basis for the hypothesis of "PJI early diagnosis and genotyping", and to lay the foundation for the study of molecular biology in the early diagnosis of PJI..

关节假体术后感染（PJI）的早期诊断是现今临床尚待解决的难题，其发病机制主要包括致病菌的黏附和生物膜的形成，早期临床标本难以收集到 “活的可培养”的致病菌，现行技术对PJI早期诊断的敏感度和特异度都不高。随着对PJI致病菌流行病学和基因分型理论研究的深入，本项目提出利用基于PJI致病菌一般和特异基因型的检测技术作为PJI早期诊断的新策略，避免了传统培养技术耗时较长且敏感度不高的缺点。选择16S rRNA作为PJI早期诊断的一般基因以提高诊断的敏感度；后续通过对PJI致病菌最常见特异基因型（ica D、eno、sar A、agr、Mec A和Mre B等基因）的组合检测以提高诊断的特异度。并通过体外和动物实验进行验证研究，如果本项目能够证实该诊断新策略具有更高的敏感度和特异度，那么将为“PJI早期诊断和基因分型”的假说提供实验依据，并能为PJI早期诊断的分子生物学理论的深入探索研究奠定基础。

关节假体术后感染（PJI）的早期诊断是现今临床尚待解决的难题，其发病机制主要包括致病菌的黏附和生物膜的形成，早期临床标本难以收集到 “活的可培养”的致病菌，现行技术对PJI早期诊断的敏感度和特异度都不高。随着对PJI致病菌流行病学和基因分型理论研究的深入，本项目提出利用基于PJI致病菌一般和特异基因型的检测技术作为PJI早期诊断的新策略，避免了传统培养技术耗时较长且敏感度不高的缺点。选择16S rRNA作为PJI早期诊断的一般基因以提高诊断的敏感度；后续通过对PJI致病菌最常见特异基因型（ica D、eno、sar A、agr、Mec A和Mre B等基因）的组合检测以提高诊断的特异度。并通过体外和动物实验进行验证研究，如果本项目能够证实该诊断新策略具有更高的敏感度和特异度，那么将为“PJI早期诊断和基因分型”的假说提供实验依据，并能为PJI早期诊断的分子生物学理论的深入探索研究奠定基础。