Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

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Delay to formalin fixation 'cold ischemia time': effect on ERBB2 detection by in-situ hybridization and immunohistochemistry.

Author(s): Portier BP, Wang Z, Downs-Kelly E, Rowe JJ, Patil D, Lanigan C, Budd GT, Hicks DG, Rimm DL, Tubbs RR

Publication: Mod Pathol, 2013, Vol. 26, Page 1-9

PubMed ID: 22899285 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of cold ischemia on the detection of ERBB2, also known as Her2, by IHC and by two methods of ISH (PathVysion FISH and the INFORM Her2 dual in situ DNA probe assay) in breast cancer tissue. The effects of using different ISH scoring criteria were also investigated. A total of 84 breast carcinoma specimens were used in this study, and IHC and ISH assays were performed using a tissue microarray.

   Summary of Findings:

   For all cold ischemia times, FISH and dual color ISH were able to detect ERBB2 amplification with 100% concordance when either the ASCO/CAP dual probe or the FDA package insert scoring methods were employed. When the ASCO/CAP single probe scoring criteria was used, FISH and dual color ISH results generally still agreed, but amplification status of 3 of the 9 amplified cases changed to equivocal or non-amplified. These 3 cases all had a cold ischemia time of <1 h. ERBB2 IHC showed the highest concordance with FISH and dual color ISH results when the single probe scoring criteria was used. Other minor discrepancies among the scoring methods and between FISH, dual color ISH, and IHC were observed, the most common being an equivocal score for IHC while FISH and dual color ISH determined a non-amplified status, regardless of scoring method. Importantly, while dual color ISH generated a stronger signal intensity than FISH for both ERBB2 and CEP17, regardless of cold ischemia time (p<0.05, all), cold ischemia times of up to 3 h did not significantly decrease signal intensities for either assay. Cold ischemia times of >3 h did however significantly decrease the FISH signal intensities for ERBB2 in stroma and chromosome 17 in stroma and tumor compared to intensities after <1 h cold ischemia time (p<0.05, all).

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Formalin

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Protein	Tissue microarray
   Protein	Immunohistochemistry
   DNA	FISH
   DNA	In situ hybridization
   DNA	Tissue microarray

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Immunohistochemistry Specific	Technology platform	Dual color in situ hybridization FISH
   FISH Specific	Data handling	ASCO/CAP dual-probe scoring ASCO/CAP single-probe scoring FDA in-situ hybridization package insert
   Biospecimen Acquisition	Cold ischemia time	<1 h 1-2 h 2-3 h >3 h
   In situ hybridization Specific	Targeted nucleic acid	ERBB2 (HER2) CEP17
   FISH Specific	Targeted nucleic acid	ERBB2 (HER2) CEP17
   In situ hybridization Specific	Detection method	Fluorescence Dual color
   Immunohistochemistry Specific	Targeted peptide/protein	ERBB2 (HER2)
   In situ hybridization Specific	Data handling	ASCO/CAP dual-probe scoring ASCO/CAP single-probe scoring FDA in-situ hybridization package insert

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