Chromogenic in-situ hybridization: a viable alternative to fluorescence in-situ hybridization in the HER2 testing algorithm

Hanna, Wedad; Kwok, Kevin W.H.

doi:10.1038/modpathol.3800555

Cited by 90 publications

(77 citation statements)

References 22 publications

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“…Some authors advocate the use of a centromeric probe on a parallel tissue section to better define the level of polysomy, particularly in borderline cases. 15 In our study, scoring based only on the EGFR signal number allows for good discrimination between low and high polysomy cases and between high polysomy and gene-amplified cases, which in most cases is easily detected as a tight cluster of signals, reflecting tandem replication at the EGFR gene locus (Po0.0001 and P ¼ 0.0003, respectively, Figure 3b and c). In the borderline cases, particularly those that appear to have between 4 and 5 signals per tumor nucleus, the parallel examination of centromeric signal may be advised, although this approach has not yet been validated.…”

Section: Discussionmentioning

confidence: 57%

“…[14][15][16] However, there is no such consensus regarding the use of EGFR in lung cancer, and published reports have adopted the HER2 approach, 23 classifying 3-5 signals per nucleus as aneuploidy, and 46 signals per nucleus in 450% of tumor cells as amplification. Our approach to CISH scoring involved evaluating 200 tumor cells per sample, recording the maximum and minimum number of signals seen in each case, and computing the geometric mean to minimize the dispersion effect of wide ranging CISH values.…”

Section: Discussionmentioning

confidence: 99%

“…The reliability of CISH as a technique for detecting gene amplification has been established for HER2 in breast cancer specimens. [14][15][16] However, there are no published reports that examine the utility of CISH in detecting EGFR copy number in lung cancer specimens. In the present study, we evaluated the utility of CISH in detecting EGFR copy number by comparing EGFR CISH results with FISH results in a cohort of 77 nonsmoking Taiwanese women with nonsmall cell lung carcinoma, and used discriminant analysis to identify CISH cut-off points that differentiate between different EGFR copy number categories.…”

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confidence: 99%

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Validation of chromogenic in situ hybridization for detection of EGFR copy number amplification in nonsmall cell lung carcinoma

et al. 2007

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Epidermal growth factor receptor (EGFR) gene copy number correlates with response to tyrosine kinase inhibitors in patients with nonsmall cell lung carcinoma. Fluorescence in situ hybridization (FISH), a standard methodology to detect EGFR copy number abnormalities in nonsmall cell lung carcinoma, is limited by instrumentation and cost. Chromogenic in situ hybridization (CISH) is an emerging alternative detection technique using light microscopy, but its utility in assessing EGFR copy number in lung cancer is not established. To address the utility of CISH, we studied paraffin-embedded nonsmall cell lung carcinoma specimens from 77 Taiwanese nonsmoking women treated by surgery alone. We recorded the number of signals per tumor cell nucleus, correlated EGFR copy number by CISH with FISH results, and used receiver operating characteristics to identify cut-off points for the CISH results. Tumors were classified as adenocarcinoma (n ¼ 28), mixed adenocarcinoma with bronchioloalveolar features (n ¼ 25), bronchioloalveolar carcinoma (n ¼ 2), squamous cell carcinoma (n ¼ 15), and adenosquamous carcinoma (n ¼ 7). By FISH, 29% of cases had no amplification, 18% had low polysomy, 35% had high polysomy, and 12% had gene amplification. EGFR copy number detected by CISH highly correlated with FISH (Spearman r ¼ 0.81, Po0.0001). We determined the optimal EGFR CISH cut-off points that discriminate between no amplification and low polysomy (2.8 signals, P ¼ 0.09); no amplification plus low polysomy and high polysomy plus gene amplification (4.5 signals, Po0.0001); and high polysomy and gene amplification (7.1 signals, P ¼ 0.0003). CISH is an alternative assay to FISH in determining EGFR copy number status that may contribute to stratification of patients with nonsmall cell lung carcinoma for clinical trials and identify a subset of patients that should be treated with tyrosine kinase inhibitors.

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Section: Discussionmentioning

confidence: 57%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Validation of chromogenic in situ hybridization for detection of EGFR copy number amplification in nonsmall cell lung carcinoma

et al. 2007

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“…A negative result is an IHC staining of 0 or 1+, a FISH result of less than 4.0 HER2 gene copies per nucleus, or FISH ratio of less than 1.8. Equivocal results require additional action for final determination (22,54) .…”

Section: Her2/neumentioning

confidence: 99%

Predictive factors of breast cancer evaluated by immunohistochemistry

Gobbi¹,

Nunes²

2008

J. Bras. Patol. Med. Lab.

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Hormone receptor and Her2 protein overexpression evaluated by immunohistochemistry (IHC) is widely validated as a predictive factor in breast cancer. The quality of the IHC reaction is influenced by tissue fixation and processing. Over-and underfixation deeply affect IHC results. Antigen retrieval may improve IHC but it does not recover tissue from autolysis or overfixation. The choice of primary antibody for IHC as to its sensitivity and specificity in relation to therapeutic response represents an important stage. Apart from mouse monoclonal antibodies, new rabbit monoclonal antibodies are commercially available, such as clones anti-ER SP1 and B644, anti-PR SP2 and B645 and anti-Her2 SP3 and 4B5. They represent an alternative to hormone receptor and Her2 evaluation by IHC. New polymeric nonbiotinylated detection systems are also available and allow accurate and strong marking with no stromal and no non-specific cytoplasmic staining due to endogenous biotin. The most recommended cut off for estrogen and progesterone receptors (ER and PR) is more than 1% of positive cells with moderate or strong staining intensity (Allred's scoring system). New guidelines for Her2 evaluation by IHC show a cut off of more than 30% of positive cells with strong intensity (3+) that correlates better with gene amplification. The 2+ cases are now considered indeterminate and should be confirmed by fluorescence in situ hybridisation (FISH)

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“…CISH has been validated in many reports compared to FISH, with a high concordance rate 6,12,18 . The current HER2 CISH is based on single-color detection, without centromere 17 correction.…”

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confidence: 99%

Rabbit antibodies for hormone receptors and HER2 evaluation in breast cancer

Nunes¹,

Sanches²,

Rocha³

et al. 2009

Rev. Assoc. Med. Bras.

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SUMMARYBACKGROUND. Novel rabbit monoclonal antibodies (RabMab) for estrogen (ER), progesterone (PR) receptors and HER2 evaluation by immunohistochemistry have recently been commercially released. We compared the RabMab anti-ER, anti-PR and anti-HER2 to mouse monoclonal antibodies (Mab) using tissue microarrays (TMA) of breast carcinomas. METHODS. Two TMA containing breast carcinomas were built. Sections were immunostained using anti-ER and anti-PR, Mab and RabMab. The sections stained for ER and PR were evaluated considering positive those tumors in which more than 1% of the tumor cell nuclei stained moderate or strong. For HER2, the immunostained sections were evaluated using the ASCO/CAP guidelines for HER2. Chromogenic in situ hybridization (CISH) was used as the gold standard for HER2 evaluation. CISH was evaluated using the Zymed HER2 CISH interpretation guidelines. RESULTS. RabMab against ER have similar staining patterns compared to the 6F11 (Mab), but stronger than 1D5 (Mab) from three different suppliers. The RabMab against PR provide stronger and sharper immunohistochemical signals compared to Mab. The detection of HER2 protein overexpression was more prevalent with the polyclonal antibodies and RabMab than with the Mab. These were more specific than the RabMab, which were more sensitive when compared to CISH. CONCLUSION. The novel RabMab against ER and PR showed higher intensity of staining than the Mab. The RabMab against HER2 is more sensitive than Mab, however, Mab presented more specificity than RabMab when compared to CISH for HER2 evaluation of breast carcinomas.

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Chromogenic in-situ hybridization: a viable alternative to fluorescence in-situ hybridization in the HER2 testing algorithm

Cited by 90 publications

References 22 publications

Validation of chromogenic in situ hybridization for detection of EGFR copy number amplification in nonsmall cell lung carcinoma

Validation of chromogenic in situ hybridization for detection of EGFR copy number amplification in nonsmall cell lung carcinoma

Predictive factors of breast cancer evaluated by immunohistochemistry

Rabbit antibodies for hormone receptors and HER2 evaluation in breast cancer

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