Røge Rasmus, Nielsen Søren, Vyberg Mogens
Institute of Pathology, Aalborg Hospital, Aarhus University Hospital.
Appl Immunohistochem Mol Morphol. 2014 Jul;22(6):449-58. doi: 10.1097/PAI.0b013e318292b764.
Immunohistochemical detection of CD15 is important in the diagnosis of Hodgkin lymphoma and may play a role in the classification of renal cell tumors (RCTs). In the NordiQC external quality assessment scheme, 4 CD15 tests, each with 71 to 121 participating laboratories, showed that 24% to 50% of the stains were insufficient. This was mainly because of very low primary antibody (Ab) concentration and insufficient heat-induced epitope retrieval, whereas the Ab clone performance seemed of little importance. The purpose of this study was to evaluate the performance of the most commonly used CD15 Abs on the basis of vendor-recommended and in-house optimized protocols. Multitissue blocks with 199 specimens including various malignant lymphomas, RCTs, and normal tissues were stained with 3 different concentrated (conc) CD15 Ab clones Carb-3, MMA, and BY87 according to predetermined in-house optimized protocols on 2 automated immunostaining platforms. Carb-3 and MMA were also applied in ready-to-use (RTU) formats utilized according to vendor protocols. Extension and intensity of stains was determined using the H-score method. Clone Carb-3-conc gave with an in-house optimized protocol the highest H-scores in Hodgkin lymphoma, RCTs, and normal kidney tissue. Clones Carb-3-RTU and MMA-conc gave slightly lower scores, whereas clones MMA-RTU and BY87-conc gave the lowest scores and a large proportion of false-negative reactions. For all concentrated Abs, in-house optimized protocols resulted in increased sensitivity and improved overall staining results compared with vendor-recommended protocols. The importance of Ab selection and protocol optimization in immunohistochemical laboratories is emphasized.
CD15的免疫组化检测在霍奇金淋巴瘤的诊断中很重要,并且可能在肾细胞肿瘤(RCT)的分类中发挥作用。在NordiQC外部质量评估计划中,4项CD15检测,每项有71至121个参与实验室,结果显示24%至50%的染色不足。这主要是由于一抗(Ab)浓度非常低以及热诱导抗原修复不足,而Ab克隆的性能似乎不太重要。本研究的目的是根据供应商推荐的方案和内部优化方案评估最常用的CD15抗体的性能。按照预定的内部优化方案,在2个自动免疫染色平台上,用3种不同浓缩度(conc)的CD15抗体克隆Carb-3、MMA和BY87对包含各种恶性淋巴瘤、RCT和正常组织的199个标本的多组织块进行染色。Carb-3和MMA也按照供应商方案以即用型(RTU)形式应用。使用H评分法确定染色的范围和强度。克隆Carb-3-conc在内部优化方案下,在霍奇金淋巴瘤、RCT和正常肾组织中给出了最高的H评分。克隆Carb-3-RTU和MMA-conc给出的分数略低,而克隆MMA-RTU和BY87-conc给出的分数最低且有很大比例的假阴性反应。对于所有浓缩抗体,与供应商推荐的方案相比,内部优化方案提高了敏感性并改善了整体染色结果。强调了免疫组化实验室中抗体选择和方案优化的重要性。