Chromogenic in situ hybridisation (CISH) should be an accepted method in the routine diagnostic evaluation of HER2 status in breast cancer

Palma, Silvia; Collins, Nadine; Faulkes, Catherine; Ping, Bruno; Ferns, Gordon A.; Haagsma, Ben; Layer, G. T.; Kissin, M W; Cook, Martin

doi:10.1136/jcp.2006.043356

Cited by 48 publications

(32 citation statements)

References 15 publications

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“…These results are consistent with other studies that have validated CISH for the determination of HER2 amplification Bhargava et al 2005;Bilous et al 2006;Di Palma et al 2007; Although FISH results are more accurate and predictive of prognosis and the response to therapy (Yaziji and Gown 2002), IHC is used as a first-line screening test. In contrast to CISH and FISH, IHC allows detection of HER2 protein levels and, therefore, directly visualises the target molecule of trastuzumab.…”

Section: Discussionsupporting

confidence: 90%

“…Chromogenic in situ hybridisation (CISH) is an attractive alternative for the detection of HER2 gene amplification, as previous studies have shown high concordance between CISH and FISH (Arnould et al 2003;Bhargava et al 2005;Bilous et al 2006;Di Palma et al 2007;Gong et al 2009a, b;Isola et al 2004;Rosa et al 2009). CISH is based on DNA probes coupled to peroxidase enzymes that catalyze chromogenic agents such as 3,3 0 -diaminobenzidine (DAB).…”

Section: Introductionmentioning

confidence: 97%

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Prospective multi-centre study to validate chromogenic in situ hybridisation for the assessment of HER2 gene amplification in specimens from adjuvant and metastatic breast cancer patients

Hoegel¹,

John

Ott

et al. 2010

J Cancer Res Clin Oncol

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 97%

Prospective multi-centre study to validate chromogenic in situ hybridisation for the assessment of HER2 gene amplification in specimens from adjuvant and metastatic breast cancer patients

Hoegel¹,

John

Ott

et al. 2010

J Cancer Res Clin Oncol

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“…11,12 The use of a light microscope instead of a fluorescence microscope and the ability to readily observe background morphologic features will be viewed as advantages compared with the current FISH assays. Based on the data from this study, the DAKO dual-color CISH assay seems to be as sensitive and specific as FISH, and, owing to its "familiarity" with immunohistochemical analysis, it may have the potential of being the preferred HER2 gene test in pathology laboratories in the future.…”

Section: Discussionmentioning

confidence: 99%

Visualization of FISH Probes by Dual-Color Chromogenic In Situ Hybridization

Hoff¹,

Jørgensen²,

Müller³

et al. 2010

Am J Clin Pathol

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A b s t r a c tThe overall purpose of the study was to demonstrate applicability of the DAKO dual-color chromogenic in situ hybridization (CISH) assay (DAKO Denmark, Glostrup) Fluorescence in situ hybridization (FISH) has achieved widespread use for a range of gene copy number detection probes and translocation probes. It is considered a very accurate and sensitive method, eg, the College of American Pathologists has published that FISH is to be regarded as a "gold standard" for HER2 testing. 1 However, the FISH method is perceived as having some limitations. The evaluation of tumor morphologic features through FISH may be difficult, and the method requires a fluorescence microscope, which is costly and not readily available in all pathology laboratories. Furthermore, the fluorescence signals fade relatively quickly, which makes archiving of the slides difficult. These limitations can be overcome by chromogenic in situ hybridization (CISH), which converts the fluorescence signals into chromogenic precipitates, and can visualize FISH-labeled probes along with the morphologic features using a brightfield microscope.We have developed a dual-color CISH assay in which the green fluorescein isothiocyanate (FITC) FISH signals are converted into blue chromogenic precipitates and the Texas red FISH signals are converted into red chromogenic precipitates. In principle, the assay can be applied on top of all FISH probes labeled with Texas red and FITC fluorochromes, including probes designed for detection of amplification and deletion in formalin-fixed, paraffin-embedded tissue sections. In relation to copy number detection, a major advantage of the dual-color approach is that the gene signals and the corresponding reference signals can be visualized in the same nuclei on 1 slide, allowing direct comparison between signals. Hence, the dualcolor technique makes it easier to distinguish true gene amplification or deletion from chromosomal aneuploidy. 2

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“…In line with the American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) guideline, the HER2 status should be determined at diagnosis for all invasive breast cancers [3]. Even though, correct assessment of the HER2 protein overexpression or gene amplification is crucial for proper treatment, studies have revealed a margin of error amounting to a mean of 25% [4,5], implying that about 10% of the patients are not treated adequately [6].…”

Section: Introductionmentioning

confidence: 99%

Digital image analysis of membrane connectivity is a robust measure of HER2 immunostains

Brügmann

Eld

Lelkaitis

et al. 2011

Breast Cancer Res Treat

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The purpose of this study was to develop and validate a new software, HER2-CONNECT(TM), for digital image analysis of the human epidermal growth factor receptor 2 (HER2) in breast cancer specimens. The software assesses immunohistochemical (IHC) staining reactions of HER2 based on an algorithm evaluating the cell membrane connectivity. The HER2-CONNECT algorithm was aligned to match digital image scorings of HER2 performed by 5 experienced assessors in a training set and confirmed in a separate validation set. The training set consisted of 167 breast carcinoma tissue core images in which the assessors individually and blinded outlined regions of interest and gave their HER2 score 0/1+/2+/3+ to the specific tumor region. The validation set consisted of 86 core images where the result of the automated image analysis software was correlated to the scores provided by the 5 assessors. HER2 fluorescence in situ hybridization (FISH) was performed on all cores and used as a reference standard. The overall agreement between the image analysis software and the digital scorings of the 5 assessors was 92.1% (Cohen's Kappa: 0.859) in the training set and 92.3% (Cohen's Kappa: 0.864) in the validation set. The image analysis sensitivity was 99.2% and specificity 100% when correlated to FISH. In conclusion, the Visiopharm HER2 IHC algorithm HER2-CONNECT(TM) can discriminate between amplified and non-amplified cases with high accuracy and diminish the equivocal category and thereby provides a promising supplementary diagnostic tool to increase consistency in HER2 assessment.

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Chromogenic in situ hybridisation (CISH) should be an accepted method in the routine diagnostic evaluation of HER2 status in breast cancer

Cited by 48 publications

References 15 publications

Prospective multi-centre study to validate chromogenic in situ hybridisation for the assessment of HER2 gene amplification in specimens from adjuvant and metastatic breast cancer patients

Prospective multi-centre study to validate chromogenic in situ hybridisation for the assessment of HER2 gene amplification in specimens from adjuvant and metastatic breast cancer patients

Visualization of FISH Probes by Dual-Color Chromogenic In Situ Hybridization

Digital image analysis of membrane connectivity is a robust measure of HER2 immunostains

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