Tafe Laura J, Allen Samantha F, Steinmetz Heather B, Dokus Betty A, Cook Leanne J, Marotti Jonathan D, Tsongalis Gregory J
Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States; Department of Pathology, Theodor and Audrey Geisel School of Medicine at Dartmouth, Hanover, NH, United States.
Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States; Department of Pathology, Theodor and Audrey Geisel School of Medicine at Dartmouth, Hanover, NH, United States.
Exp Mol Pathol. 2014 Aug;97(1):116-9. doi: 10.1016/j.yexmp.2014.06.003. Epub 2014 Jun 11.
HER2 fluorescence in-situ hybridization (FISH) is used in breast and gastro-esophageal carcinoma for determining HER2 gene amplification and patients' eligibility for HER2 targeted therapeutics. Traditional manual processing of the FISH slides is labor intensive because of multiple steps that require hands on manipulation of the slides and specifically timed intervals between steps. This highly manual processing also introduces inter-run and inter-operator variability that may affect the quality of the FISH result. Therefore, we sought to incorporate an automated processing instrument into our FISH workflow.
Twenty-six cases including breast (20) and gastro-esophageal (6) cancer comprising 23 biopsies and three excision specimens were tested for HER2 FISH (Pathvysion, Abbott) using the Thermobrite Elite (TBE) system (Leica). Up to 12 slides can be run simultaneously. All cases were previously tested by the Pathvysion HER2 FISH assay with manual preparation. Twenty cells were counted by two observers for each case; five cases were tested on three separate runs by different operators to evaluate the precision and inter-operator variability.
There was 100% concordance in the scoring between the manual and TBE methods as well as among the five cases that were tested on three runs. Only one case failed due to poor probe hybridization. In total, seven cases were positive for HER2 amplification (HER2:CEP17 ratio >2.2) and the remaining 19 were negative (HER2:CEP17 ratio <1.8) utilizing the 2007 ASCO/CAP scoring criteria. Due to the automated denaturation and hybridization, for each run, there was a reduction in labor of 3.5h which could then be dedicated to other lab functions.
The TBE is a walk away pre- and post-hybridization system that automates FISH slide processing, improves work flow and consistency and saves approximately 3.5h of technologist time. The instrument has a small footprint thus occupying minimal counter space. TBE processed slides performed exceptionally well in comparison to the manual technique with no disagreement in HER2 amplification status.
人表皮生长因子受体2（HER2）荧光原位杂交（FISH）技术用于乳腺癌和胃食管癌的检测，以确定HER2基因扩增情况及患者是否适合接受HER2靶向治疗。传统的FISH载玻片手工处理过程劳动强度大，因为有多个步骤需要手动操作载玻片，且各步骤之间有特定的时间间隔。这种高度依赖手工的处理方式还会引入批间和操作者间的差异，可能影响FISH结果的质量。因此，我们试图将一种自动化处理仪器纳入我们的FISH工作流程。
使用徕卡公司的Thermobrite Elite（TBE）系统，对26例病例（包括20例乳腺癌和6例胃食管癌，共23份活检标本和3份切除标本）进行HER2 FISH（Pathvysion，雅培公司）检测。该系统最多可同时处理12张载玻片。所有病例之前均通过Pathvysion HER2 FISH检测法进行过手工制片检测。两名观察者对每个病例计数20个细胞；5例病例由不同操作者在三个不同批次进行检测，以评估其精密度和操作者间的差异。
手工方法与TBE方法之间以及在三个批次检测的5例病例之间的评分一致性为100%。仅1例因探针杂交不佳而失败。根据2007年美国临床肿瘤学会/美国病理学家协会（ASCO/CAP）的评分标准，总共7例病例HER2扩增呈阳性（HER2:CEP17比值>2.2），其余19例为阴性（HER2:CEP17比值<1.8）。由于采用了自动变性和杂交技术，每次检测可减少3.5小时的人工，这些时间可用于其他实验室工作。
TBE是一种杂交前后无需人工操作的系统，可实现FISH载玻片处理的自动化，改善工作流程和一致性，节省技术人员约3.5小时的时间。该仪器占地面积小，占用的台面空间最小。与手工技术相比，TBE处理的载玻片表现出色，HER2扩增状态的判定无差异。
