Bright-field In Situ Hybridization for HER2 Gene Amplification in Breast Cancer Using Tissue Microarrays Correlation Between Chromogenic (CISH) and Automated Silver-enhanced (SISH) Methods With Patient Outcome
Bright-field In Situ Hybridization for HER2 Gene Amplification in Breast Cancer Using Tissue Microarrays Correlation Between Chromogenic (CISH) and Automated Silver-enhanced (SISH) Methods With Patient Outcome
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DOI：
10.1097/pdm.0b013e31816f6374
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发表时间：
2009-06-01
影响因子：
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通讯作者：
Stein, Sandra R.
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文献类型：
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作者：
Francis, Glenn D.;Jones, Mark A.;Stein, Sandra R.
前言：HER2基因扩增或过度表达在乳腺癌中的发生率为15%~25%，对乳腺癌的治疗和预后有重要意义。HER2检测最常用的方法是荧光原位杂交(FISH)和免疫组织化学。FISH被认为是参考标准，可以更准确地预测曲妥珠单抗的疗效，但技术要求高，价格昂贵，需要专门的设备。在澳大利亚，要求原位杂交才有资格接受曲妥珠单抗的辅助治疗。明场原位杂交是FISH的全部替代方法，使用原位方法和过氧化物酶介导的显色底物的组合，如二氨基联苯胺[显色原位杂交(CISH)]或多聚体技术与酶金相[银增强原位杂交(SISH)]相结合来创建在明场显微镜下可见的标记。2006年10月，在澳大利亚的诊断检测中引入了《国际清洁卫生标准》。SISH方法是最近引入测试系统的一种方法。采用组织微阵列技术对593例浸润性乳腺癌患者的临床资料进行组织微阵列的构建，并对其结果进行比较。使用美国临床肿瘤学会/美国病理学家学会指南和澳大利亚HER2咨询委员会标准(单探针：二倍体，1-2.5拷贝/核；多倍体；2.5-4拷贝/核；可疑，4-6拷贝/核；低水平扩增，6-10拷贝/核和高水平扩增>；10拷贝/核；双探针HER2/CHR17比率：未扩增；1.8，不明确1.8-2.2，扩增>；结果：337个组织核心，230个患者样本的结果是有信息的。HER2单探针对CHISH和SISH的符合率为96%，单探针对HER2/CHR17SISH的符合率为95.5%。结论：HER2 SISH检测结合了自动化和明视场显微镜的优点，方便了实验室内的工作流程，缩短了周转时间，并与患者的预后相关。
Introduction: HER2 gene amplification or overexpression occurs in 15% to 25%, of breast cancers and has implications for treatment and prognosis. The most commonly used methods for HER2 testing are fluorescence in situ hybridization (FISH) and immunohistochemistry. FISH is considered to be the reference standard and more accurately predicts response to trastuzumab, but is technically demanding, expensive, and requires specialized equipment. In situ hybridization is required to be eligible for adjuvant treatment with trastuzumab in Australia. Bright-field in situ hybridization is all alternative to FISH and uses a combination of in situ methodology and a peroxidase-mediated chromogenic Substrate such as diaminobenzidine [chromogenic in situ hybridization (CISH)] or multimer technology coupled with enzyme metallography [silver-enhanced in situ hybridization (SISH)] to create a marker visible under bright-field microscopy. CISH was introduced into diagnostic testing in Australia in October 2006. SISH methodology is a more recent introduction into the testing repertoire. An evaluation of CISH and SISH performance to assess patient outcome were performed using tissue microarrays.Materials and Methods: Tissue microarrays were constructed in duplicate using material front 593 patients with invasive breast carcinoma and assessed using CISH and SISH. Gene amplification was assessed using the American Society of Clinical Oncology/College of American Pathologists guideline and Australian HER2 Advisory Board criteria (single probe: diploid, 1 to 2.5 copies/nucleus; polysomy > 2.5 to 4 copies/nucleus; equivocal, > 4 to 6 copies/nucleus; low-level amplification, > 6 to 10 copies/nucleus and high-level amplification > 10 copies/nucleus; dual probe HER2/CHR17 ratio: nonamplified < 1.8, equivocal 1.8 to 2.2, amplified > 2.2).Results: Results were informative for 337 tissue cores comprising 230 patient samples. Concordance rates were 96% for HER2 single probe CISH and SISH and 95.5% for single probe CISH and dual probe HER2/CHR17 SISH. Both bright-field methods correlated with immunohistochemistry results and with breast cancer-specific survival.Conclusions: HER2 SISH testing combines the advantages of automation and bright-field microscopy to facilitate workflow within the laboratory, improves turnaround time, and correlates with patient outcome.