Pathology and Laboratory Medicine Institute, Cleveland Clinic, Cleveland, OH, USA.
Mod Pathol. 2013 Jan;26(1):1-9. doi: 10.1038/modpathol.2012.123. Epub 2012 Aug 17.
The American Society of Clinical Oncology/College of American Pathologists ERBB2 testing guidelines address several pre-analytical variables known to affect ERBB2 testing accuracy. According to 2010 updated guidelines, the pre-analytical variable of time to tissue fixation (cold ischemia time) should be kept to <1 h, however, little has been published about cold ischemia time and its significance in ERBB2 testing. To that end, this study evaluated ERBB2 status using two different FDA-approved in-situ hybridization methods and an FDA-approved immunohistochemistry (IHC) assay in the largest cohort to date (n=84) of invasive breast carcinomas with tracked cold ischemia time. Cold ischemia time was stratified into four groups (<1 h (n=45), 1-2 h (n=27), 2-3 h (n=6), and >3 h (n=6)) and ERBB2 status was evaluated in each group by IHC (4B5) and by in-situ hybridization methodologies (PathVysion(®) fluorescence in situ hybridization and the INFORM HER2(®) dual in situ DNA probe assay). Both in-situ hybridization methods were evaluated using three ERBB2 scoring criteria (dual-probe guidelines, single-probe guidelines, and the FDA package insert scoring instructions). Fluorescence in-situ hybridization (FISH) and INFORM HER2(®) demonstrated 100% concordance in the detection of ERBB2 amplification by all three scoring guidelines at all cold ischemia time points. Agreement between in-situ hybridization methodologies and IHC was superior using single-probe guidelines compared with dual probe or FDA scoring instructions. In addition, Inform HER2(®) in-situ hybridization signals were significantly more intense than FISH at all cold ischemia time points, however, no significant loss of either chromosome 17 or ERBB2 signal was detected by FISH or Inform HER2(®) in-situ hybridization in cold ischemia times up to 3 h. On the basis of our findings, cold ischemia time up to 3 h has no deleterious effect on the detection of ERBB2 via in-situ hybridization or IHC.
美国临床肿瘤学会/美国病理学家协会 ERBB2 检测指南涉及多个已知会影响 ERBB2 检测准确性的分析前变量。根据 2010 年更新的指南，组织固定（冷缺血时间）的分析前变量应保持在<1 小时内，然而，关于冷缺血时间及其在 ERBB2 检测中的意义，相关研究发表较少。为此，本研究使用两种不同的美国食品药品监督管理局批准的原位杂交方法和一种美国食品药品监督管理局批准的免疫组织化学（IHC）检测方法，对迄今为止最大的（n=84）具有跟踪冷缺血时间的浸润性乳腺癌队列评估 ERBB2 状态。将冷缺血时间分为四组（<1 小时（n=45）、1-2 小时（n=27）、2-3 小时（n=6）和>3 小时（n=6）），并在每个组中通过 IHC（4B5）和原位杂交方法（PathVysion（®）荧光原位杂交和 INFORM HER2（®）双原位 DNA 探针检测）评估 ERBB2 状态。两种原位杂交方法均使用三种 ERBB2 评分标准（双探针指南、单探针指南和美国食品药品监督管理局试剂盒说明书评分说明）进行评估。荧光原位杂交（FISH）和 INFORM HER2（®）在所有冷缺血时间点均通过所有三种评分标准检测 ERBB2 扩增，显示出 100%的一致性。与双探针或美国食品药品监督管理局评分说明相比，使用单探针指南，原位杂交方法与 IHC 的一致性更高。此外，在所有冷缺血时间点，Inform HER2（®）原位杂交信号均比 FISH 更强烈，但在冷缺血时间长达 3 小时内，FISH 或 Inform HER2（®）原位杂交均未检测到染色体 17 或 ERBB2 信号的明显丢失。基于我们的研究结果，冷缺血时间长达 3 小时对通过原位杂交或 IHC 检测 ERBB2 没有有害影响。
