Limits...
Out of the darkness and into the light: bright field in situ hybridisation for delineation of ERBB2 (HER2) status in breast carcinoma.

Gruver AM, Peerwani Z, Tubbs RR - J. Clin. Pathol. (2010)

Bottom Line: However, neither of these modalities is without limitations.The average percentage agreement in an informal analysis of studies comparing HER2 amplification by chromogenic in situ hybridisation with FISH was 96% (SD 4%); kappa coefficients ranged from 0.76 to 1.0.Although a much smaller number of studies are available for review, similar levels of concordance have been reported in studies comparing HER2 amplification by methods employing metallography (silver in situ hybridisation) with FISH.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pathology, Pathology and Laboratory Medicine Institute, Cleveland Clinic, Lerner College of Medicine, Cleveland, Ohio 44195, USA.

ABSTRACT
Assessment of ERBB2 (HER2) status in breast carcinomas has become critical in determining response to the humanised monoclonal antibody trastuzumab. The current joint College of American Pathologists and the American Society of Clinical Oncology guidelines for the evaluation of HER2 status in breast carcinoma involve testing by immunohistochemistry and fluorescence in situ hybridisation (FISH). However, neither of these modalities is without limitations. Novel bright field in situ hybridisation techniques continue to provide viable alternatives to FISH testing. While these techniques are not limited to evaluation of the HER2 gene, the extensive number of studies comparing bright field in situ techniques with other methods of assessing HER2 status allow a robust evaluation of this approach. Analysis of the literature demonstrates that, when used to assess HER2 gene status, bright field in situ hybridisation demonstrates excellent concordance with FISH results. The average percentage agreement in an informal analysis of studies comparing HER2 amplification by chromogenic in situ hybridisation with FISH was 96% (SD 4%); kappa coefficients ranged from 0.76 to 1.0. Although a much smaller number of studies are available for review, similar levels of concordance have been reported in studies comparing HER2 amplification by methods employing metallography (silver in situ hybridisation) with FISH. A summary of the advancements in bright field in situ hybridisation, with focus on those techniques with clinical applications of interest to the practicing pathologist, is presented.

Show MeSH

Related in: MedlinePlus

Conceptual schematic demonstrating dual detection of HER2 and chromosome 17 by bright field double in situ hybridisation. By this technique, dual detection can be accomplished using individual single haptens. The two probes are incompatible and two rounds of target DNA denaturation, hybridisation and stringency washes are carried out sequentially. DNP, dinitrophenol.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC2921277&req=5

fig5: Conceptual schematic demonstrating dual detection of HER2 and chromosome 17 by bright field double in situ hybridisation. By this technique, dual detection can be accomplished using individual single haptens. The two probes are incompatible and two rounds of target DNA denaturation, hybridisation and stringency washes are carried out sequentially. DNP, dinitrophenol.

Mentions: Dual-colour FISH is considered the ‘gold standard’ for in situ assessment of gene copy number, in part because of the superior spatial resolution offered by this technique and the FDA approval status of Vysis PathVysion.12 However, dual colour FISH has the same disadvantages as single-colour FISH and the additional limitation that probes producing more intense signal may lead to the interpretation of biased ratios favouring the brighter probe. Despite some limitations, the ability to directly assess both and multiple targets in the same nucleus simultaneously is highly desirable. Although studies evaluating multicolour detection procedures for bright field microscopy using chromosome specific probes had been reported in the past,80–82 development of dual-colour CISH using probes for HER2 and chromosome 17 was reported more recently using single-colour detection of a digoxigenin-labelled HER2 probe and a biotin labelled chromosome 17 probe.83 The results of dual-coloured CISH and FISH in that study showed high concordance (91%, κ coefficient 0.82), and the contrast provided by the two colours allowed for immediate distinction between HER2 amplification and chromosome 17 aneuploidy.84 Additional reports of dual-colour CISH for the assessment of HER2 gene status found excellent concordance when respectively compared with FISH results (98.6% and 94.6%).67 85 Additional advancements in bright field in situ hybridisation are aiming to provide assessment of both HER2 and chromosome 17 through techniques to identify both targets either simultaneously or consecutively (figure 5). Examples of the staining produced by such techniques are demonstrated in figure 6.


Out of the darkness and into the light: bright field in situ hybridisation for delineation of ERBB2 (HER2) status in breast carcinoma.

Gruver AM, Peerwani Z, Tubbs RR - J. Clin. Pathol. (2010)

Conceptual schematic demonstrating dual detection of HER2 and chromosome 17 by bright field double in situ hybridisation. By this technique, dual detection can be accomplished using individual single haptens. The two probes are incompatible and two rounds of target DNA denaturation, hybridisation and stringency washes are carried out sequentially. DNP, dinitrophenol.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2921277&req=5

fig5: Conceptual schematic demonstrating dual detection of HER2 and chromosome 17 by bright field double in situ hybridisation. By this technique, dual detection can be accomplished using individual single haptens. The two probes are incompatible and two rounds of target DNA denaturation, hybridisation and stringency washes are carried out sequentially. DNP, dinitrophenol.
Mentions: Dual-colour FISH is considered the ‘gold standard’ for in situ assessment of gene copy number, in part because of the superior spatial resolution offered by this technique and the FDA approval status of Vysis PathVysion.12 However, dual colour FISH has the same disadvantages as single-colour FISH and the additional limitation that probes producing more intense signal may lead to the interpretation of biased ratios favouring the brighter probe. Despite some limitations, the ability to directly assess both and multiple targets in the same nucleus simultaneously is highly desirable. Although studies evaluating multicolour detection procedures for bright field microscopy using chromosome specific probes had been reported in the past,80–82 development of dual-colour CISH using probes for HER2 and chromosome 17 was reported more recently using single-colour detection of a digoxigenin-labelled HER2 probe and a biotin labelled chromosome 17 probe.83 The results of dual-coloured CISH and FISH in that study showed high concordance (91%, κ coefficient 0.82), and the contrast provided by the two colours allowed for immediate distinction between HER2 amplification and chromosome 17 aneuploidy.84 Additional reports of dual-colour CISH for the assessment of HER2 gene status found excellent concordance when respectively compared with FISH results (98.6% and 94.6%).67 85 Additional advancements in bright field in situ hybridisation are aiming to provide assessment of both HER2 and chromosome 17 through techniques to identify both targets either simultaneously or consecutively (figure 5). Examples of the staining produced by such techniques are demonstrated in figure 6.

Bottom Line: However, neither of these modalities is without limitations.The average percentage agreement in an informal analysis of studies comparing HER2 amplification by chromogenic in situ hybridisation with FISH was 96% (SD 4%); kappa coefficients ranged from 0.76 to 1.0.Although a much smaller number of studies are available for review, similar levels of concordance have been reported in studies comparing HER2 amplification by methods employing metallography (silver in situ hybridisation) with FISH.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pathology, Pathology and Laboratory Medicine Institute, Cleveland Clinic, Lerner College of Medicine, Cleveland, Ohio 44195, USA.

ABSTRACT
Assessment of ERBB2 (HER2) status in breast carcinomas has become critical in determining response to the humanised monoclonal antibody trastuzumab. The current joint College of American Pathologists and the American Society of Clinical Oncology guidelines for the evaluation of HER2 status in breast carcinoma involve testing by immunohistochemistry and fluorescence in situ hybridisation (FISH). However, neither of these modalities is without limitations. Novel bright field in situ hybridisation techniques continue to provide viable alternatives to FISH testing. While these techniques are not limited to evaluation of the HER2 gene, the extensive number of studies comparing bright field in situ techniques with other methods of assessing HER2 status allow a robust evaluation of this approach. Analysis of the literature demonstrates that, when used to assess HER2 gene status, bright field in situ hybridisation demonstrates excellent concordance with FISH results. The average percentage agreement in an informal analysis of studies comparing HER2 amplification by chromogenic in situ hybridisation with FISH was 96% (SD 4%); kappa coefficients ranged from 0.76 to 1.0. Although a much smaller number of studies are available for review, similar levels of concordance have been reported in studies comparing HER2 amplification by methods employing metallography (silver in situ hybridisation) with FISH. A summary of the advancements in bright field in situ hybridisation, with focus on those techniques with clinical applications of interest to the practicing pathologist, is presented.

Show MeSH
Related in: MedlinePlus