Comparison of molecular diagnosis with serum markers and synovial fluid analysis in patients with prosthetic joint infection
Comparison of molecular diagnosis with serum markers and synovial fluid analysis in patients with prosthetic joint infection
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DOI：
10.1302/0301-620x.100b10.bjj-2018-0096.r1
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发表时间：
2018-10-01
影响因子：
4.6
通讯作者：
Lee, M. S.
中科院分区：
文献类型：
--
作者：
Kuo, F-C;Lu, Y-D;Lee, M. S.
目的本研究的目的是比较16 S/28 S rRNA测序结果与红细胞沉降率（ESR）、C反应蛋白（CRP）水平和滑液分析在诊断人工关节感染（PJI）中的作用。在研究人群中，有25名PJI患者和189名对照。在PJI患者中，14例（56%）为女性，诊断时的平均年龄为65岁（38至83岁）。检测ESR和CRP水平，并前瞻性采集滑液标本。对滑液进行靶向16 S/28 S rRNA的逆转录聚合酶链反应（RT-PCR）/序列分析，并进行常规培养。对临床信息不知情的实验室人员进行了所有检测。PJI的诊断是根据肌肉骨骼感染Society.ResultsA共25例确诊PJI的标准。在20例单菌性PJI中，PCR产物与序列分析所提供的不同菌种的16 S/28 SrRNA基因完全匹配。在5例PJI的多菌病例中，16 S/28 SrRNAPCR序列分析未能鉴定出一致的细菌种类。在189名对照患者中，有一个假阳性RT-PCR结果。分子诊断方法的敏感性和特异性均为100%（95%置信区间（CI）85.7至100）和99.5%（95% CI 97.1 - 99.9），而PCR的阳性和阴性预测值分别为96.1%（95%CI 79.6 ~ 99.9）和100%（95%CI 98.1 ~ 100）。PCR检测结果明显优于血清学诊断方法（ESR和CRP分别为p = 0.004和p = 0.010），滑液白色血细胞（WBC）计数（p = 0.036），和多形核白细胞百分比（PMN%）结论16 S/16 S引物的逐步RT-PCR和序列分析可用于检测人乳腺癌组织中的16 S/16 S融合蛋白。在严格的实验室条件下进行的28 S rRNA在区分无菌和脓毒症关节方面获得了高度敏感和特异的结果接受关节成形术序列分析成功地鉴定了单一微生物感染中的细菌菌株，但未能鉴定多微生物感染中的分子靶标。需要进一步完善鉴定多种微生物感染中细菌的方案。
AimsThe aim of this study was to compare the results of 16S/28S rRNA sequencing with the erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) level, and synovial fluid analysis in the diagnosis of prosthetic joint infection (PJI).Patients and MethodsBetween September 2015 and August 2016, 214 consecutive patients were enrolled. In the study population, there were 25 patients with a PJI and 189 controls. Of the PJI patients, 14 (56%) were women, and the mean age at the time of diagnosis was 65 years (38 to 83). The ESR and CRP levels were measured, and synovial fluid specimens were collected prospectively. Synovial fluid was subjected to reverse transcription polymerase chain reaction (RT-PCR)/sequence analysis targeting the 16S/28S rRNA, and to conventional culture. Laboratory personnel who were blind to the clinical information performed all tests. The diagnosis of PJI was based on the criteria of the Musculoskeletal Infection Society.ResultsA total of 25 patients had a confirmed PJI. In 20 cases of monomicrobial PJI, the PCR products could be perfectly matched with the 16S/28S rRNA genes specific for different species of bacteria provided by sequence analysis. Of the five polymicrobial cases of PJI, 16S/28S rRNA PCR sequence analysis failed to identify the concordant bacteria species. In the 189 control patients, there was one false-positive RT-PCR result. The sensitivity and specificity of the molecular diagnosis method were 100% (95% confidence interval (CI) 85.7 to 100) and 99.5% (95% CI 97.1 to 99.9), respectively, whereas the positive and negative predictive values of PCR were 96.1% (95% CI 79.6 to 99.9) and 100% (95% CI 98.1 to 100), respectively. The PCR results were significantly better than serological diagnostic methods (p = 0.004 and p = 0.010 for ESR and CRP, respectively), the synovial fluid white blood cell (WBC) count (p = 0.036), and percentage of polymorphonuclear cells (PMN%) (p = 0.014).ConclusionStepwise RT-PCR and sequence analysis of the 16S/28S rRNA carried out under stringent laboratory conditions achieved highly sensitive and specific results for the differentiation between aseptic and septic joints undergoing arthroplasty. Sequence analysis successfully identified bacterial strains in monomicrobial infections but failed to identify molecular targets in polymicrobial infections. Further refinement of the protocols to identify the bacteria in polymicrobial infections is needed.