Kölbel B, Wienert S, Dimitriadis J, Kendoff D, Gehrke T, Huber M, Frommelt L, Tiemann A, Saeger K, Krenn V
Zentrum für Histologie, Zytologie und Molekulare Diagnostik, Max-Planck-Str. 5, 54296, Trier, Deutschland.
Z Rheumatol. 2015 Sep;74(7):622-30. doi: 10.1007/s00393-015-1571-8.
The aim of this project was to devise a quantification method for neutrophils within a single focal point through the development of a CD15 focus score which enables bacterial infections in synovial-like interface membranes (SLIM) to be diagnosed.
In this study a histopathological classification of 91 SLIM removed during revision surgery from the hips (n = 59) and knees (n = 32) was performed. Neutrophils were identified immunohistochemically by means of a CD15-specific monoclonal antibody. The quantitative evaluation of CD15-positive neutrophils (CD15Ne) used the principle of maximum focal infiltration (focus) together with an assessment of a single focal point (0.3 mm(2)). This immunohistochemical approach made it possible to develop the CD15 quantifier software, which automatically quantifies CD15Ne.
The SLIM cases with positive microbiological findings (n = 47) had significantly (p < 0.001, Mann-Whitney U-test) more CD15Ne/focal point than cases with negative microbiological findings (n = 44). A count of 50 CD15Ne/focal point was identified as the optimum threshold when diagnosing periprosthetic joint infections (PJI) using the CD15 focus score. If the microbiological findings are used as a gold standard, the diagnostic sensitivity is 0.83, and the specificity is 0.864 with a positive predictive value (PPV) of 0.87, a negative predictive value (NPV) of 0.83, an accuracy of 0.846 and an area under the curve (AUC) of 0.878. The evaluation of findings for the preparations using the CD15 quantifier software (n = 31) deviated by an average of 12 cells from the histopathological evaluation findings (CD15 focus score). Above a cell count of 62, the CD15-quantifier needs on average 32 s less than the pathologist.
The immunohistochemical CD15 focus score has a high diagnostic value and allowed the development of the CD15 quantifier software. This provides an automated procedure, which shortens the mentally tiring and time-consuming process of microscopic cell counting and thus makes a contribution towards the standardization of tools for diagnosing PJI.
本项目的目的是通过开发CD15焦点评分来设计一种针对单个焦点内中性粒细胞的量化方法,从而能够诊断滑膜样界面膜(SLIM)中的细菌感染。
在本研究中,对91例在翻修手术中从髋关节(n = 59)和膝关节(n = 32)取出的SLIM进行了组织病理学分类。通过CD15特异性单克隆抗体免疫组化鉴定中性粒细胞。对CD15阳性中性粒细胞(CD15Ne)的定量评估采用最大焦点浸润(焦点)原则以及对单个焦点(0.3平方毫米)的评估。这种免疫组化方法使得开发CD15定量软件成为可能,该软件可自动对CD15Ne进行定量。
微生物学检查结果为阳性的SLIM病例(n = 47),其每个焦点的CD15Ne数量显著(p < 0.001,曼-惠特尼U检验)多于微生物学检查结果为阴性的病例(n = 44)。当使用CD15焦点评分诊断假体周围关节感染(PJI)时,每个焦点50个CD15Ne的计数被确定为最佳阈值。如果将微生物学检查结果作为金标准,诊断敏感性为0.83,特异性为0.864,阳性预测值(PPV)为0.87,阴性预测值(NPV)为0.83,准确性为0.846,曲线下面积(AUC)为0.878。使用CD15定量软件对制剂结果进行评估(n = 31),与组织病理学评估结果(CD15焦点评分)平均相差12个细胞。当细胞计数超过62时,CD15定量软件平均比病理学家少用32秒。
免疫组化CD15焦点评分具有较高的诊断价值,并促成了CD15定量软件的开发。这提供了一种自动化程序,缩短了显微镜下细胞计数这一令人精神疲惫且耗时的过程,从而有助于实现PJI诊断工具的标准化。