Sompuram Seshi R, Vani Kodela, Tracey Brian, Kamstock Debra A, Bogen Steven A
Medical Discovery Partners LLC, Boston, Massachusetts (SRS,KV,SAB)
Department of Electrical and Computer Engineering, Tufts University, Medford, Massachusetts (BT)
J Histochem Cytochem. 2015 Sep;63(9):681-90. doi: 10.1369/0022155415588109. Epub 2015 May 4.
A new standardized immunohistochemistry (IHC) control for breast cancer testing comprises formalin-fixed human epidermal growth factor receptor 2, estrogen receptor, or progesterone receptor peptide antigens covalently attached to 8-µm glass beads. The antigen-coated beads are suspended in a liquid matrix that hardens upon pipetting onto a glass microscope slide. The antigen-coated beads remain in place through deparaffinization, antigen retrieval, and immunostaining. The intensity of the beads' stain provides feedback regarding the efficacy of both antigen retrieval and immunostaining. As a first report, we tested the sensitivity and specificity of the new IHC controls ("IHControls"). To evaluate sensitivity, various staining problems were simulated. IHControls detected primary and secondary reagent degradation similarly to tissue controls. This first group of IHControls behaved similarly to tissue controls expressing high concentrations of the antigen. The IHControls were also able to detect aberrations in antigen retrieval, as simulated by sub-optimal times or temperatures. Specificity testing revealed that each antigen-coated bead was specific for its cognate IHC test antibody. The data support the conclusion that, like tissue controls, IHControls are capable of verifying the analytic components of an immunohistochemical stain. Unlike tissue controls, IHControls are prepared in large bulk lots, fostering day-to-day reproducibility that can be standardized across laboratories.
一种用于乳腺癌检测的新型标准化免疫组织化学（IHC）对照物，由共价连接到8微米玻璃珠上的福尔马林固定的人表皮生长因子受体2、雌激素受体或孕激素受体肽抗原组成。包被抗原的珠子悬浮在一种液体基质中，当用移液管移到玻璃显微镜载玻片上时，该基质会硬化。包被抗原的珠子在脱石蜡、抗原修复和免疫染色过程中保持原位。珠子染色的强度提供了关于抗原修复和免疫染色效果的反馈。作为首次报告，我们测试了新型IHC对照物（“IH对照物”）的敏感性和特异性。为了评估敏感性，模拟了各种染色问题。IH对照物检测一级和二级试剂降解的情况与组织对照物相似。这第一组IH对照物的表现与表达高浓度抗原的组织对照物相似。IH对照物还能够检测抗原修复过程中的异常情况，如通过次优时间或温度模拟的情况。特异性测试表明，每个包被抗原的珠子对其对应的IHC测试抗体具有特异性。数据支持这样的结论：与组织对照物一样，IH对照物能够验证免疫组织化学染色的分析成分。与组织对照物不同，IH对照物是大批量制备的，促进了日常的可重复性，并且可以在各实验室之间进行标准化。
