Immunoperoxidase versus immunofluorescence in the assessment of human renal biopsies
Immunoperoxidase versus immunofluorescence in the assessment of human renal biopsies
复制标题
DOI：
10.1053/j.ajkd.2004.12.019
复制
发表时间：
2005-04-01
影响因子：
13.2
通讯作者：
Svalander, CT
中科院分区：
文献类型：
--
作者：
Mölne, J;Breimer, ME;Svalander, CT
背景世纪来，冰冻切片免疫荧光（IF）一直是肾活检标本免疫组化评价的金标准。在常规诊断免疫组织病理学评价中，传统的IF已在很大程度上被应用于甲醛固定组织的石蜡切片的免疫过氧化物酶（IP）方法所取代。这部分是由于IF方法固有的实际缺点造成的，例如，分离的组织标本和处理、UV显微镜、标签制作存档的褪色和不持久以及后期调查困难。本研究的目的是评价IP作为IF的替代品在肾活检标本诊断评估中的作用。研究方法：将蛋白水解抗原修复、对脱蜡切片有效的抗体、灵敏的检测系统（Dako EnVision HRP; Dako，哥本哈根，丹麦）以及标准化和严格控制的程序应用于先前通过光学显微镜（LM）和IF分类的一系列肾活检标本（n = 81）。假设IF为试验标准品，将免疫球蛋白G（IgG）、伊加、IgM、C1 q和C3 c染色按配对比例记录为IF和IP阳性或阴性。结果如下：所有观察结果的一致性为71%（282/398次观察），IgG为82%（65/79次观察），伊加为89%（72/81次观察）。大多数不一致的观察结果（116个观察结果中的74个）为阳性，其中113个为阳性，系膜区IgM和C1 q沉积物未被IF发现。统计学上，IF和IP之间IgG、伊加和C3 c的结局无显著差异（P > 0.2）。此外，IP染色允许通过LM同时评价组织，因此组织结构和免疫沉积物之间的相关性不容易通过IF获得。结论：在本研究中，记录了对于IgG、伊加和C3 c的检测，除少数例外情况外，IP应用于蛋白酶消化的去甲氨蝶呤固定的肾组织脱蜡切片与冷冻切片上的IF相同。此处使用的EnVision HRP方法在一抗稀释方面比早期现有的IP方法有效数倍，并且由于不涉及抗生物素蛋白-生物素系统，因此很少会发生非特异性背景染色。由于IgM和C1 q的阳性IP结果，大多数（91/116次观察）观察结果不一致（116/398次观察; 29%），值得进一步研究。(c)2005年，美国国家肾脏基金会（National Kidney Foundation，Inc.）
Background For half a century, immunofluorescence (IF) on frozen sections has been the gold standard for immunohistochemical evaluation of renal biopsy specimens. In routine diagnostic immunohistopathologic evaluation, traditional IF has been replaced to a large extent by immunoperoxidase (IP) methods applied to paraffin sections of formaldehyde-fixed tissue. This is caused in part by the practical disadvantages inherent in the IF method, eg, separate tissue specimen and handling, UV microscopy, fading and impermanence of the label-making archiving, and difficult later investigation. Our aim for the present study is to evaluate IP as an alternative to IF in the diagnostic assessment of renal biopsy specimens. Methods: Proteolytic antigen retrieval, antibodies effective on deparaffinized sections, a sensitive detection system (Dako EnVision HRP; Dako, Copenhagen, Denmark), and a standardized and rigorously controlled procedure were applied to a series of renal biopsy specimens (n = 81) previously classified by means of light microscopy (LM) and IF. Staining for immunoglobulin G (IgG), IgA, IgM, C1q, and C3c were recorded as positive or negative for IF and IP in paired proportions, presuming that IF was the test standard. Results: Concordant observations were 71% for all (282 of 398 observations), 82% for IgG (65 of 79 observations), and 89% for IgA (72 of 81 observations). The majority of discordant observations (74 of 116 observations) were positive by means of 113, with mesangial deposits of IgM and C1q that were not found by IF. Statistically, there was no significant difference in outcomes between IF and IP for IgG, IgA, and C3c (P > 0.2). In addition, IP staining allowed simultaneous evaluation of tissue by LM and therefore correlation between tissue structure and immune deposits not readily attained by IF. Conclusion: In the present study, it is documented that for the detection of IgG, IgA, and C3c, IP applied to protease-digested deparaffinized sections of formaldehyde-fixed renal tissue is, with few exceptions, equal to IF on frozen sections. The EnVision HRP method used here is several times more effective in terms of primary antibody dilution than earlier existing IP methods, and because the avidin-biotin system is not involved, very little nonspecific background staining will occur. Discordant observations (116 of 398 observations; 29%) were in the majority (91 of 116 observations) due to positive IP findings of IgM and C1q, which deserve additional investigation. (c) 2005 by the National Kidney Foundation, Inc.