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Test Code IHC for Semi-Quant HER2 Immunohistochemistry for Semi-Quantitative HER2

Important Note

Order as "HMISC"

Useful For

Determining over-expression of HER2 protein in formalin-fixed, paraffin-embedded tissue sections. Tissues frequently evaluated for HER2 overexpression are breast, gastric, and esophageal invasive cancers.

Synonyms/Keywords

Histology, HER2/Neu, Surgical Pathology, Immunohistochemistry, C-erbB2, HER2, Gastric, Breast, Esophageal, IHC, Immunohistochemistry for Semi-Quantitative HER2, HMISC

Specimen Requirements

Specimen Type Preferred Container/Tube Acceptable Container/Tube Specimen Volume Specimen Minimum Volume
(allows for 1 repeat)		 Pediatric Minimum Volume
(no repeat)
Paraffin- Embedded Tissue Block or Unstained Sections on Charged Slides   Transport Container for Block and/or Slide Holders      
Completed Pathology Report          

Acceptable Body Sites

Breast. gastric, and/or their metastases

Acceptable Specimen Types

Paraffin-embedded tissue blocks and/or unstained charged slides fixed in 10% neutral buffered formalin.  Unstained slides require an H&E slide or an additional unstained slide to be stained for H&E.

Collection/Processing Instructions

A. BREAST

1. Ischemic Time - removal to initiation of fixation be less than or equal to one hour.
2. Fixative type - 10% Neutral Buffered Formalin.
3. Fixed in 10% NBF for at least 6 hours and less than 72 hours

The above handling and fixative times must be documented on the accession slip or the report or both according to ASCO/CAP guidelines as delayed, under, or over fixation may affect test results.

 

Handling of Surgical Breast Specimens Obtained Remotely
The following procedure, for breast specimens when cancer is suspected/known. The goal is to expose breast tumor to formalin within one hour of removal.

  • Breast lumpectomy
  • Pat the specimen dry
  • Ink the margins - Inking: Unoriented specimens are inked one color. Oriented specimens are inked six colors. You can ink however you want but write on the requisition slip/histology order how you have inked it. One suggestion for inking:         
    • Superior - blue         
    • Inferior - green         
    • Medial - red         
    • Lateral - orange         
    • Anterior/Superficial - yellow         
    • Posterior/Deep - black
  • Dry again and dip in vinegar (helps the ink stick)
  • Make an incision in the specimen as close as possible to the lesion. Do not cut all the way through the specimen - keep it intact. More than one incision can be made on larger specimens.
  • Cut opposite the margin closest to the tumor.  

 

Remember the specimen needs to be inked before it is incised in order to maintain integrity of the margin. Inking after the margin has been cut is not recommended. Specimen should be immersed in a sufficient volume of 10% neutral buffered formalin (10X volume of specimen).


Requirements:

  • Completed histology requisition form or Authorization for Test Requests form.
  • Surgical or cytology pathology report containing pertinent patient history, tissue source, and diagnosis.
  • One paraffin tissue/cell block with corresponding H&E slide, otherwise send 2 (4-5 micron) unstained, air-dried, charged slides with corresponding H&E slide.  

 

NOTE: For HER2 FISH testing, send 2 (4-5 micron) unstained, air-dried, charged slides with corresponding H & E slide.     

 

Contact Histology Pathology Assistants for more information  

 

B. GASTRIC/ESOPHAGEAL RESECTIONS OR BIOPSY
Fixative type - 10% Neutral Buffered Formalin. It is recommended (not required) that since proper fixation is critical for quality staining, resection and biopsy specimens be handled similar to that of breast specimens.  

 

Requirements:

  • Completed histology requisition form or Authorization for Test Requests form and include: Surgical or cytology pathology report containing pertinent patient history, tissue source, and diagnosis.
  • One paraffin tissue/cell block with corresponding H&E slide, otherwise send 2 (4-5 micron) unstained, air-dried, charged slides with corresponding H&E slide.  

 

NOTE: For HER2 FISH testing, send 2 (4-5 micron) unstained, air-dried, charged slides with corresponding H & E slide.   

 

HER2 FISH testing is a send out test.   

 

Antibody validation was performed on 10% neutral buffered, formalin-fixed, paraffin embedded tissues/cell blocks.

Specimen Stability Information

Specimen Type Temperature Time
Paraffin Tissue/Cell Block, Fixed in 10% NBF Room Temperature/Ambient Indefinite
Unstained, Air-Dried, Charged Slides Room Temperature/Ambient Ideally slides should be ≤ to 6 weeks old

Rejection Criteria

No Invasive Cancer Present
Fixative other than 10% Neutral Buffered Formalin
Decalcified Tissue (if necessary, will require a disclaimer).
Unstained Slides (no charge on slides)
Insufficient Data (Incomplete requisition and/or Pathology Report) may delay testing.

Interferences

  • Proper fixation conditions are critical for quality staining. Under or over fixation, or delay to fixative can affect quality results. These tests are not validated on fixatives other than 10% neutral buffered formalin or decalcified tissues.

Interpretations

Test is only available with interpretation.  

 

Interpretation: Semi-Quantitative Immunohistochemistry, Manual  

 

FISH HER 2 may be ordered and is available separately. All 2+ results will automatically be tested for FISH Her2 at an additional fee.  

 

HER2 FISH testing is a send out test.

 

All blocks and slides will be returned to the requestor.


1. BREAST CANCER Requirements

  • Time to fixation ≤ 1hr
  • Fixed in 10% Neutral Buffered Formalin (NBF)
  • Fixed in 10% NBF for at least 6 hours and less than 72 hours

HER2/NEU Scoring System and Criteria
 

IHC Result Criteria
0 Negative No staining is observed* or membrane staining that is incomplete and is faint/barely perceptible and within 10% of invasive tumor cells.
1+ Negative Incomplete membrane staining that is faint/barely perceptible and within >10% of invasive tumor cells*.
2+ Equivocal Weak to moderate complete membrane staining observed in >10% of tumor cells.
3+ Positive Circumferential membrane staining that is complete, intense, and within >10% of invasive tumor cells*.
 * See comment Indeterminate

Technical issues prevent tests from being reported as positive, negative, or equivocal, which may include:         

  • Inadequate specimen handling         
  • Artifacts that make interpretation difficult         
  • Analytic testing failure         
  • Sample has strong membrane staining of normal breast ducts

Over-expression of HER2 cell membrane receptor protein in breast carcinoma is associated with tumor cell growth, aggressive disease and shortened survival. A positive test result aids in the assessment for possible treatment with Herceptin (trastuzumab) that is targeted against the HER2 receptor protein. The prognostic and therapeutic significance of the results of this assay in other tumors is under investigation. The FDA has approved the reagents used in this immunohistochemistry assay for assessment of HER2 receptor status. The range of over-expression among breast cancers is reportedly between 10% to 20% of primary breast carcinomas.

 

2. GASTRIC/ESOPHAGEAL RESECTION Requirements
Fixed in 10% Neutral Buffered Formalin (NBF)


HER2/NEU Scoring System and Criteria

IHC Result Criteria
0 Negative No reactivity or membranous reactivity in < 10% of tumor cells
1+ Negative Faint/barely perceptible membranous reactivity in ≥ 10% of tumor cells; cells reactive only in part of their membrane (Reflex to FISH)
2+ Equivocal Weak to moderate complete, basolateral or lateral membranous reactivity in ≥ 10% of tumor cells
3+ Positive Strong, complete, basolateral or lateral membranous reactivity in ≥10% of tumor cells

 

3. GASTRIC/ESOPHAGEAL BIOPSY Requirements

  • Fixed in 10% Neutral Buffered Formalin (NBF)

 

HER2/NEU Scoring System and Criteria

IHC Result Criteria
0 Negative No reactivity in any tumor cells
1+ Negative Tumor cell cluster (≥5 cells) with a faint/barely perceptible membranous reactivity irrespective of tumor cells stained
2+ Equivocal Tumor cell cluster (≥5 cells) with weak to moderate complete, basolateral or lateral membranous reactivity irrespective of percentage of tumor cells stained (Reflex to FISH)
3+ Positive Tumor cell cluster (≥5 cells) with a strong complete, basolateral or lateral membranous reactivity irrespective of percentage of tumor cells stained.

Depending on the series, the prevalence of HER2 expression in gastric cancer seems to be 15-25%. The level of expression of intestinal type gastric cancer seems to be much higher (~32%) compared with diffuse-type gastric cancer (~6%). Also, tumors that are primarily located at the gastroesophageal junction seem to have higher HER2 positive rates compared to tumors that occur in the rest of the stomach (33% versus 21%).(7)  

 

ALL
The clone used is a mouse monoclonal, CB11 and detection system used is a polymer based. This test is validated on paraffin-embedded, 10% neutral formalin fixed tiossues.  Interpretation follows the American Society for Clinical Oncology (ASCO) and College of American Pathologists (CAP) guidelines. 

 

References:

1. Reference: Wolff AC, Hammond ME, Allison KH, Harvey BE, Mangu PB, Bartlett JM, Bilous M, Ellis IO, Fitzgibbons P, Hanna W, Jenkins RB, Press MF, Spears PA, 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;36:210522.

2. Walker Ra, Bartlett JMS Dowsett M, Ellis IO, Hanby An, Jasani B, Miller K, and Pinder SE.  HER2 Testing in the UK-Further Update to Recommendations.  Journal of Clinical Pathology 2008. 

3. Yaziji H, Goldstein LC, Barry TS, Werling R, Hwang H, Ellis GK, Gralow JR, Livingston RB, Gown AM: HER-2 testing in breast cancer using parallel tissue-based methods. JAMA 2004, 291:1972-1977.

4.Owens MA, Horten BC, Da Silva MM: HER2 amplification ratios by fluorescence in situ hybridization and correlation with immunohistochemistry in a cohort of 6556 breast cancer tissues. Clin Breast Cancer 2004, 5:63-69.

5. Hofmann M et al, Assessment of a HER2 scoring system for gastric cancer: results from a validation study. Histopathol 52:797. 2008.

6. Bang Y et al, Pathologic features of advanced gastric cancer (GC): relationship to human epidermal growth factor receptor 2 (HER2) positivity in the global screening programme of the ToGA trial. J Clin Oncol 25:15s (suppl; abstr 4556).

(7) Reference:

Reference Information: Questions Relating to Immunohistochemistry for HER2 on Gastric and Gastroesophageal Junction Carcinomas

Reference Range Information

Performing Location Reference Range
Marshfield Interpretative Report Semi-Quantitative/IHC 

Classification

This test was developed and its performance characteristics determined by Marshfield Labs. It has not been cleared or approved by the U.S. Food and Drug Administration. The FDA has determined that such clearance or approval is not necessary. This test is for clinical purposes. It should not be regarded as investigational or for research. This laboratory is certified under the Clinical Laboratory Improvement Amendments (CLIA) as qualified to perform high complexity testing.    

Marshfield Labs Performing Department

Marshfield Labs Histology

Performing Information

Performing Location Day(s) Test Performed Analytical Time Methodology/Instrumentation
Marshfield Monday through Friday 36 hours Polymer based detection system/Leica Bond III

CPT Codes

CPT Modifier
(if needed)		 Quantity Description Comments
88360   1    

Outreach CPT Codes

CPT Modifier
(if needed)		 Quantity Description Comments
88360   1    

Ordering Applications

Ordering Application Description
Cerner Her-2 (c-erbB2) Oncoprotein