Test Catalog

Test Id : H2BR

HER2 Amplification Associated with Breast Cancer, FISH, Tissue

Useful For
Suggests clinical disorders or settings where the test may be helpful

A predictive marker for patients with both node-positive or node-negative primary and metastatic breast cancer

 

Patients with HER2 amplification that may be candidates for therapies targeting the human epidermal growth factor receptor 2 (HER2) protein (eg, trastuzumab [Herceptin], pertuzumab, lapatinib)

 

Confirming the presence of HER2 amplification in cases with 2+ (low level) or 3+ (high level) HER2 overexpression by immunohistochemistry, and for certain histologic subtypes with aberrant patterns of HER2 expression seen by immunohistochemistry (eg, micropapillary carcinoma)

Reflex Tests
Lists tests that may or may not be performed, at an additional charge, depending on the result and interpretation of the initial tests.

Test Id Reporting Name Available Separately Always Performed
HERBN HER Breast IHC Automated NO Reflex Yes No

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

Reflex testing will be performed using immunohistochemistry (IHC) when the fluorescence in situ hybridization (FISH) result falls within certain ranges as defined by the 2018 focused update to the American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines.(1) For FISH results in ASCO/CAP categories Group 2, 3, and 4 (formerly called "equivocal"), the IHC testing will be added, charged, and reported separately. An integrated interpretation of the IHC and FISH results will be provided (see Interpretation).

 

Initial results are typically completed within 6 days. If results of the initial probe set indicate IHC reflex testing is necessary based on ASCO/CAP guidelines, complete results will typically be available within 8 days.

 

A charge and CPT code is applied for each probe set hybridized, analyzed, and reported.

Method Name
A short description of the method used to perform the test

Fluorescence In Situ Hybridization (FISH)

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.

Yes

Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

HER2, Breast Tumor, FISH, Tissue

Aliases
Lists additional common names for a test, as an aid in searching

Breast Carcinoma

c-erb-b2 Amplification Test (FISH)

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

Reflex testing will be performed using immunohistochemistry (IHC) when the fluorescence in situ hybridization (FISH) result falls within certain ranges as defined by the 2018 focused update to the American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines.(1) For FISH results in ASCO/CAP categories Group 2, 3, and 4 (formerly called "equivocal"), the IHC testing will be added, charged, and reported separately. An integrated interpretation of the IHC and FISH results will be provided (see Interpretation).

 

Initial results are typically completed within 6 days. If results of the initial probe set indicate IHC reflex testing is necessary based on ASCO/CAP guidelines, complete results will typically be available within 8 days.

 

A charge and CPT code is applied for each probe set hybridized, analyzed, and reported.

Specimen Type
Describes the specimen type validated for testing

Tissue

Ordering Guidance

This test is only for primary or metastatic breast tumors.

-For urothelial tumors, order H2UR / HER2 Amplification Associated with Urothelial Carcinoma, FISH, Tissue.

-For gastroesophageal tumors, order H2GE / HER2 Amplification Associated with Gastroesophageal Cancer, FISH, Tissue.

-For all other tumor types, order H2MT / HER2 Amplification, Miscellaneous Tumor, FISH, Tissue.

 

This test does not include a pathology consult. If a pathology consult is requested, PATHC / Pathology Consultation should be ordered and the appropriate fluorescence in situ hybridization test will be determined by the reviewing pathologist and testing performed at an additional charge.

Shipping Instructions

Advise Express Mail or equivalent if not on courier service.

Necessary Information

1. A pathology report is required in order for testing to be performed. Acceptable pathology reports include working drafts, preliminary pathology, or surgical pathology reports.

2. A reason for testing must be provided. If this information is not provided, an appropriate indication for testing may be entered by Mayo Clinic Laboratories.

3. The pathology report must include type and time of fixation, as well as the cold ischemia time.

Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

Note: In accordance to College of American Pathologists guidelines, place specimens for HER2 (ERBB2) testing in fixative within one hour of biopsy or resection (cold ischemia time). Specimens should remain in 10% neutral buffered formalin for a minimum of six hours to a maximum of 72 hours (formalin fixation time). Do not use decalcification solutions with strong acids.(2)

 

Submit only 1 of the following specimens:

 

Specimen Type: Tissue

Preferred: Tissue block

Collection Instructions: Submit a formalin-fixed, paraffin-embedded (FFPE) tumor tissue block.

 

Acceptable: Slides

Collection Instructions: Four consecutive, unstained, 5 micron-thick sections placed on positively charged slides, and 1 hematoxylin and eosin-stained slide.

Forms

If not ordering electronically, complete, print, and send a Oncology Test Request (T729) with the specimen.

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the Testing Laboratory

Two consecutive, unstained, 5-micron-thick sections placed on positively charged slides, and 1 hematoxylin and eosin-stained slide.

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

All specimens will be evaluated at Mayo Clinic Laboratories for test suitability.

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

Specimen Type Temperature Time Special Container
Tissue Ambient (preferred)
Refrigerated

Useful For
Suggests clinical disorders or settings where the test may be helpful

A predictive marker for patients with both node-positive or node-negative primary and metastatic breast cancer

 

Patients with HER2 amplification that may be candidates for therapies targeting the human epidermal growth factor receptor 2 (HER2) protein (eg, trastuzumab [Herceptin], pertuzumab, lapatinib)

 

Confirming the presence of HER2 amplification in cases with 2+ (low level) or 3+ (high level) HER2 overexpression by immunohistochemistry, and for certain histologic subtypes with aberrant patterns of HER2 expression seen by immunohistochemistry (eg, micropapillary carcinoma)

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

Reflex testing will be performed using immunohistochemistry (IHC) when the fluorescence in situ hybridization (FISH) result falls within certain ranges as defined by the 2018 focused update to the American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines.(1) For FISH results in ASCO/CAP categories Group 2, 3, and 4 (formerly called "equivocal"), the IHC testing will be added, charged, and reported separately. An integrated interpretation of the IHC and FISH results will be provided (see Interpretation).

 

Initial results are typically completed within 6 days. If results of the initial probe set indicate IHC reflex testing is necessary based on ASCO/CAP guidelines, complete results will typically be available within 8 days.

 

A charge and CPT code is applied for each probe set hybridized, analyzed, and reported.

Clinical Information
Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

HER2 (ERBB2: c-erb-b2) is an oncogene on the long arm of chromosome 17 that is amplified in approximately 15% to 20% of breast cancers. Amplification or overexpression of HER2 has been shown to be associated with shorter disease-free survival and poorer overall survival in breast cancers. Patients with HER2 gene amplification or overexpression are candidates for treatment with the drugs that target the human epidermal growth factor receptor 2 (HER2) protein or its downstream pathways (eg, trastuzumab [Herceptin], pertuzumab).

 

Fluorescence in situ hybridization with labeled DNA probes to the pericentromeric region of chromosome 17 and to the HER2 locus can be used to determine if a patient's breast cancer has HER2 gene amplification. Immunohistochemical analysis is used to determine if a tumor exhibits HER2 overexpression.

Reference Values
Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.

An interpretive report will be provided.

Interpretation
Provides information to assist in interpretation of the test results

An interpretive report will be provided. Results are interpreted utilizing the current American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines.(1)

 

Under the 2018 Focused Update to the ASCO/CAP Guidelines, reflex immunohistochemistry (IHC) is performed for certain categories of results, known as Groups 2, 3, and 4. These categories are shown in the table below (Group 4 is the category formerly referred to as fluorescence in situ hybridization (FISH) "equivocal"). If reflex IHC is performed and is either negative (0, 1+) or positive (3+), the result of the FISH assay is considered resolved by IHC as either negative or positive. If the IHC assay shows an equivocal (2+) result, then the FISH slide is re-scored within the areas showing the most intense membranous (2+) staining and the final FISH result is used to determine whether the result is negative or positive.

 

ASCO/CAP result category

HER2:D17Z1 ratio;

average HER2 copies per cell

Reporting approach per 2018 ASCO/CAP guidelines

Group 1

HER2:D17Z1 =2.00; HER2/cell > or =4.0

Positive

Group 2

HER2:D17Z1 =2.00; HER2/cell <4.0

Reflex IHC; FISH re-analysis if 2+

Group 3

HER2:D17Z1 <2.00; HER2/cell > or =6.0

Reflex IHC; FISH re-analysis if 2+

Group 4

HER2:D17Z1 <2.00; HER2/cell > or =4.0 <6.0

Reflex IHC; FISH re-analysis if 2+

Group 5

HER2:D17Z1 <2.00; HER2/cell <4.0

Negative

 

The degree of HER2 amplification varies in tumors. Some exhibit high levels of amplification (HER2:D17Z1 ratio >4.0), whereas others exhibit low-level amplification (HER2:D17Z1 ratio of 2.0-4.0). It is not currently known if patients with different levels of amplification have the same prognosis and response to therapy.

 

Reports also interpret the HER2 copy number changes relative to chromosome 17 copy number (aneusomy) or potential structural genomic abnormalities that increase HER2 copy number.

 

Rare cases may not show HER2 amplification but still have human epidermal growth factor receptor 2 (HER2) protein overexpression demonstrated by immunohistochemistry. The clinical significance of HER2 protein overexpression in the absence of HER2 gene amplification is unclear. However, these patients may have a worse prognosis and be candidates for treatments that target the HER2 protein or its downstream pathways.

Cautions
Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

Optimum fixation should be between 6 and 72 hours in 10% neutral buffered formalin. Other types of fixatives should not be used.

 

The prognostic information provided by the HER2 status of a patient's tumor should not be interpreted in isolation because other prognostic features (eg, lymph node status, tumor size, estrogen/progesterone receptor status) may be of equal or greater importance in determining the patient's prognosis.

Supportive Data

The probe was independently validated in a blinded study on 1156 paraffin-embedded breast tissue samples. The results of the fluorescence in situ hybridization testing was correlated to the immunohistochemical analysis, which was interpreted on a scale ranging from 0 to 3+ according to FDA-approved guidelines.

Clinical Reference
Recommendations for in-depth reading of a clinical nature

1. Wolff AC, Hammond MEH, Allison KH, et al: Human Epidermal Growth Factor Receptor 2 Testing in Breast Cancer: American Society of Clinical Oncology/College of American Pathologists Clinical Practice Guideline Focused Update. J Clin Oncol. 2018 Jul 10;36(20):2105-2122 doi:10.1200/JCO.2018.77.8738

2. College of American Pathologists Cytogenetics Checklist item CYG.48932. 2019 Sep:25

3. Wolff AC, Hammond ME, Hicks DG, et al: Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society for Clinical Oncology/College of American Pathologists clinical practice guideline update. J Clin Onc. 2013 Nov 1;31(31):3997-4013

4. Perez EA, Roche PC, Jenkins RB, et al: HER2 testing in patients with breast cancer: poor correlation between weak positively by immunohistochemistry and gene amplification by fluorescence in situ hybridization. Mayo Clin Proc. 2002 Feb;77(2):148-154

5. Romond EH, Perez EA, Bryant J, et al: Trastuzumab plus adjuvant chemotherapy for operable HER2-positive breast cancer. N Engl J Med. 2005 Oct 20;353(16):1673-1684

6. Perez EA, Romond EH, Suman VJ, et al: Four-year follow-up of trastuzumab plus adjuvant chemotherapy for operable human epidermal growth factor receptor 2-positive breast cancer: joint analysis of data from NCCTG N9831 and NSABP B-31. J Clin Oncol. 2011 Sep 1;29(25):3366-3373

7. Blumenthal GM, Scher NS, Cortazar P, et al: First FDA approval of dual anti-HER2 regimen: pertuzumab in combination with trastuzumab and docetaxel for HER2-positive metastatic breast cancer. Clin Cancer Res. 2013 Sep 15;19(18):4911-4916

8. Robidoux A, Tang G, Rastogi P: Lapatinib as a component of neoadjuvant therapy for HER2-positive operable breast cancer (NSABP protocol B-41): an open-label, randomized phase 3 trial. Lancet Oncol. 2013 Nov;14(12):1183-1192

Method Description
Describes how the test is performed and provides a method-specific reference

The test is performed using the PathVysion HER2 DNA probe set (Abbott Molecular) with a HER2 probe and a chromosome 17 centromere probe (D17Z1). Paraffin-embedded tissues are cut at 5 microns and mounted on positively charged glass slides. The selection of tissue and the identification of target areas on the hematoxylin and eosin (H and E)-stained slide are performed by a pathologist. Using the H and E-stained slide as a reference, target areas are etched with a diamond-tipped etcher on the back of the unstained slide to be assayed. The probe is hybridized to the appropriate target areas and 2 technologists each analyze 30 interphase nuclei (60 total) with the results expressed as a ratio HER2:D17Z1 signals. The results are interpreted based on the 2018 guidelines established by the American Society of Clinical Oncology (ASCO) and College of American Pathologists (CAP).(1)

 

Reflex testing using immunohistochemistry will be performed when the HER2 FISH result is Group 2, Group 3, or Group 4 (based on 2018 Focused Update to ASCO/CAP guidelines).(Unpublished Mayo method)

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information

No

Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Monday through Friday

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

6 to 8 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

Slides and H&E used for analysis are retained by the laboratory in accordance to CAP and NYS requirements. Client provided paraffin blocks and extra unstained slides (if provided) will be returned after testing is complete.

Performing Laboratory Location
Indicates the location of the laboratory that performs the test

Rochester

Fees
Several factors determine the fee charged to perform a test. Contact your U.S. or International Regional Manager for information about establishing a fee schedule or to learn more about resources to optimize test selection.

  • Authorized users can sign in to Test Prices for detailed fee information.
  • Clients without access to Test Prices can contact Customer Service 24 hours a day, seven days a week.
  • Prospective clients should contact their Regional Manager. For assistance, contact Customer Service.

Test Classification
Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) product.

This test was developed, and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the US Food and Drug Administration.

CPT Code Information
Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.

88377

 

LOINC® Information
Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the order and results codes of this test. LOINC values are provided by the performing laboratory.

Test Id Test Order Name Order LOINC Value
H2BR HER2, Breast Tumor, FISH, Tissue 96893-3
Result Id Test Result Name Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
603074 Result Summary 50397-9
603075 Interpretation 69965-2
603076 Result 62356-1
GC028 Reason for Referral 42349-1
603077 Specimen 31208-2
603078 Source 85303-6
603079 Tissue ID 80398-1
603080 Fixative 8100-0
603081 Method 85069-3
603082 Additional Information 48767-8
603083 Disclaimer 62364-5
603084 Released By 18771-6

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

Excel | Create a PDF

Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports