Additional PathVysion information

PATHVYSION HER-2 DNA PROBE KIT

 

Reliable Results

The PathVysion HER-2 DNA probe kit is one of the first examples of what is recognized as genomic disease management, or personalized medicine. This means that the test helps enable the accurate assessment of a patient's HER-2 status at the DNA level with a high degree of accuracy and helps guide doctors to make the most appropriate therapy decisions based on the patient's own genetic profile.

 

PathVysion is one of the preferred tests for assessing HER-2 status:

  • FISH is considered the gold standard for HER-2 testing.
  • Studies indicate FISH to accurately and reproducibly assess tumors for HER-2 status.
  • PathVysion exhibits better concordance than IHC between local and central laboratories
  • The Chromosome 17 probe acts as an internal control and corrects for polysomy (identifies gene amplification versus chromosome 17 polysomy) and nuclear truncation
  • FDA approved automated result scanning is available for PathVysion**

** http://www.fda.gov/ Abbott AutoVysion approved Oct., 2004, Bioview Duet approved Oct. 2008, Ikonisys oncoFISH her2 approved Oct., 2008

 

PathVysion direct labeled FISH probes yield definitive results at the molecular level. The test uses fluorescent probes to "paint" the HER-2 genes in the tumor cell nucleus, to see if the number of gene copies is normal or not. Interpretation is objective - results are obtained by counting the fluorescent signals (dots). In addition, because this test measures genetic material, which is very stable, tissue preparation has very little effect on test outcome.

 
 
 
 
Patients

Welcome

A woman newly diagnosed with breast cancer should work with her doctor to determine the next steps for both testing and treatment. HER-2 status provides information that can help you and your doctor decide how to best approach your therapy choices.

patient-home.jpg

This section provides the information you need to know:

About HER-2 and HER-2 Testing

HER-2

HER-2 stands for Human Epidermal Growth Factor Receptor 2. The HER-2 gene is one of thousands of genes that comprise every living person's genetic code and is present in the nucleus of all human cells, including breast tissue cells. The HER-2 gene provides the blueprint for the manufacture of the HER-2 protein. This protein, which is attached to the cell membrane, regulates normal cell growth and function.

 

Abnormally high quantities of the HER-2 gene have been associated with rapid tumor cell growth, resistance to therapy, and shorter disease-free periods and overall survival. Approximately 20-25%, or 1 out of every 4, breast cancers have an abnormally high quantity of the HER-2 gene, and are considered HER-2 positive. 

 

HER-2 Testing

Doctors now recognize that knowing a patient's HER-2 gene status is important in understanding how to treat breast cancer.

 

HER-2 status is a strong, independent predictor of prognosis, and disease free progression. Certain breast cancer treatments are more effective for treating women who have an excess of the HER-2 gene. Therefore, determining a woman's HER-2 status can represent a critical point for the doctor to decide how to approach a woman's therapy choices. HER-2 positive patients may respond to an anti-HER-2 therapy called Herceptin® (trastuzumab), a targeted therapy for treating HER-2 positive breast cancer.

 

The risks of incorrect HER-2 status assessment can be significant: false positive assessments may lead to inappropriate medical decisions, causing a patient to undergo unnecessary therapy. Conversely, a false negative assessment may deprive a patient of potentially beneficial therapy.

 

For more information on Herceptin®, visit www.herceptin.com.

 

Who Should be Tested?

Both the National Comprehensive Cancer Network (NCCN®) and the American Society of Clinical Oncology (ASCO®) recommend determining HER-2 status for all patients with invasive breast cancer.

 

While it is recommended at initial diagnosis, HER-2 status can be determined at any time after initial diagnosis as well. There is discussion within the medical community about whether HER-2 status should be assessed with each occurrence or recurrence of invasive breast cancer.

 

Types of HER-2 Testing

HER-2 Testing Options—Why FISH technology is important

Tests for determining HER-2 status measure either gene copy number or protein. The two most widely used types of tests are ImmunoHistoChemistry (IHC) and Fluorescence in situHybridization (FISH). The PathVysion test employs FISH technology to determine HER-2 status.

 

The IHC test is a protein-based test that is used to determine the total amount of HER-2 protein receptors on the surface of the cell. To do this, the surface of the cell is "stained" with an antibody. The surface protein measured by IHC can be damaged during preparation of the tissue sample and this may cause variability in outcome. A pathologist must judge the degree of color change in the cell to determine a HER-2 protein measurement level of 0, 1+, 2+, or 3+. Because this test is relatively subjective, the results often are confirmed by performing a FISH test.

 

FISH is a gene-based test that measures the number of HER-2 genes in a cell. The FISH test "highlights" the HER-2 genes inside the cell, making them appear fluorescent so they may be accurately counted. If a patient has more than two genes per cell, she is considered HER-2 positive. If the test shows a normal gene count, the test results are considered HER-2 negative. FISH appears to be an alternative technique capable of overcoming many of the inherent technical and interpretative limitations of other techniques, such as immunohistochemistry.

 

Why PathVysion May be Right for You

The PathVysion HER-2 DNA Probe Kit can be a valuable tool used in the fight against breast cancer and provides physicians, pathologists and patients with highly accurate and reliable HER-2 results.

 

The PathVysion HER-2 test is one of the first examples of what is recognized as genomic disease management, or personalized medicine. This means that the test enables the assessment of a patient's HER-2 status at the DNA level with a high degree of accuracy and helps guide doctors to make the most appropriate therapy decisions based on a patient's own genetic profile.

 

PathVysion is approved by the U.S Food and Drug Administration (FDA) for use as an aid in determining prognostic factors in patients with stage II, node-positive breast cancer, and as an aid to predict disease-free and overall survival in patients with stage II, node-positive breast cancer treated with adriamycin-based chemotherapy and Herceptin®(Trastuzumab) monoclonal antibody therapy selection. It is not intended for use in screening for or diagnosing breast cancer.

 

PathVysion has been proven to be a highly accurate and reliable test for assessing HER-2 status:

 

  • FISH is considered the gold standard for HER-2 testing.
  • Studies indicate that FISH provides more accurate and more reproducible assessment for tumor's HER-2 status.

 

Talking With Your Doctor

To help doctors give their patients the best possible care, the American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) partnered with medical experts to develop recommendations for improving the accuracy of HER-2 testing for patients with invasive breast cancer. As a result, ASCO/CAP issued a joint clinical practice guideline about HER-2 testing for breast cancer.

 

To learn more about these guidelines, please visit Cancer.net online to review the ASCO Patient Guide: HER-2 Testing for Breast Cancer.

 

Questions To Ask Your Doctor

  • The following list of questions will help you start a meaningful dialogue with your doctor.
  • What type of breast cancer do I have?
  • Can you give me information about HER-2 testing?
  • Have I been tested for HER-2 status?
  • Is the PathVysion FISH test right for me?
 
 
 
clinicians

Welcome

Advances in breast cancer testing and treatment have greatly improved patient outcomes. Knowing a woman's HER-2 status is an essential component to understanding how to treat breast cancer and potentially extend survival.

 

This webpage will provide you with an overview of the HER-2 testing options currently available and why studies have consistently shown that the PathVysion HER-2 test, which employs FISH technology, can help overcome technical and interpretative limitations of other testing methods to accurately and reproducibly assess a woman's tumor for HER-2 status.

 

This section provides the information you need to know:

 

The Expanding Role of HER-2 Testing

HER-2 status can be a powerful tool, along with existing clinical and pathologic information, to assist in predicting prognosis and disease free progression. Accurate assessment of HER-2 status can be important in selecting appropriate therapy, including Herceptin® therapy.

 

Herceptin® (trastuzumab) is now approved in the adjuvant setting to treat early-stage HER-2 positive breast cancer, and is approved as both a first-line therapy in combination with chemotherapy and as a single agent in second- and third-line therapy.

 

The risks of incorrect HER-2 status assessment can be significant: false positive assessments may lead to inappropriate medical decisions, causing a patient to undergo unnecessary therapy. Conversely, a false negative assessment may deprive a patient of potentially beneficial therapy.

 

HER-2 Testing Options

An accurate HER-2 assessment can enable the most appropriate therapy decision.

 

Tests for HER-2 status measure either gene copy number or presence of protein receptors. The two most widely used technologies worldwide and approved by the FDA in the United States are Immunohistochemistry (IHC) and Fluorescence in situ Hybridization (FISH). Chromogenic in situ Hybridization (CISH) and Silver in situ Hybridization (SISH) are two other technologies that measure HER-2 gene copy number. All of these techniques are routinely utilized on formalin-fixed, paraffin-embedded breast cancer tissue.

 

Florescent in situ Hybridization (FISH)

FISH is a cytogenetic technique that can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes. It uses fluorescent probes that can be visualized with a fluorescent microscope.

 

FISH Process

fishprocess5.jpg

 

FISH for HER-2 is a gene-based test that measures the number of HER-2 genes in a cell. The FISH test "highlights" the HER-2 genes inside the cell, making them appear as fluorescent signals (dots) so they may be accurately counted. PathVysion® FISH also measures the number of copies of chromosome 17 in the cell. Since the HER-2 gene resides on chromosome 17, this adds several measures of control to the test. After counting the HER-2 and Chromosome 17 signals in 20 nuclei, the ratio of HER-2 to Chromosome 17 is calculated. If this value is 2 or greater, the patient is considered amplified for HER-2 or positive. If the test shows a normal gene count, she is considered HER-2 negative. FISH allows the viewer to literally count the genes, which can help overcome technical and interpretative limitations of other testing methods. In addition, because this test measures genetic material, which is very stable, tissue preparation has very little effect on test outcome.

 

her2-pathimage.gif

Images are examples of PathVysion results. Individual results may vary.

 

ImmunoHistoChemistry (IHC)

IHC is a technique that utilizes specific antibodies for staining protein in tissues on microscopic glass slides. Immunohistochemical staining is widely used in the diagnosis and treatement of many types of cancer.

 

The IHC test for HER-2 is a protein-based test that is used to determine the total amount of HER-2 protein receptors on the surface of the cell. The surface of the cell is "stained" with an antibody. IHC is interpreted by the intensity and percentage of cells which exhibit a brown cell surface staining. These specimens are interpreted on a numeric scale of 0 (negative for HER-2 overexpression), 1+ (negative for HER-2 overexpression), 2+ (positive for HER-2 overexpression), and 3+ (positive for HER-2 overexpression).

 

Issues regarding the accuracy of this method include:

  • The protein may be damaged by the processes of fixing and storing the tissue with formalin and parrafin, causing variability in test outcome;
  • An assortment of antibodies are used for the testing, including laboratory developed antibodies, which can lead to variability; and 
  • Interpretation is relatively subjective and requires a great level of skill and expertise.

ihc-sampleimages.gif

Images are examples of IHC results. Individual results may vary. 

 

Why PathVysion May Be Right For Your Patients

 

The PathVysion HER-2 DNA Probe Kit can be a valuable tool in the fight against breast cancer. It is a precise test that provides clinicians and their patients with accurate and reliable HER-2 results.

 

The PathVysion HER-2 DNA probe kit is one of the first examples of what is recognized as genomic disease management, or personalized medicine - meaning that the test enables the accurate assessment of a patient's HER-2 status at the DNA level and guides doctors to make the most appropriate therapy decisions based on the patient's own genetic profile.

 

PathVysion is approved by the U.S Food and Drug Administration (FDA) for use as an aid in determining prognostic factors in patients with stage II, node-positive breast cancer, as an aid to predict disease-free and overall survival in patients with stage II, node-positive breast cancer treated with adriamycin-based chemotherapy, and assisting in Herceptin® (Trastuzumab) monoclonal antibody therapy selection.

 

PathVysion is one of the preferred methods to asses HER-2 status:

  • FISH is considered the gold standard for HER-2 testing
  • Studies indicate PathVysion can help overcome technical and interpretative limitations of other testing methods, including IHC.

PathVysion direct labeled FISH probes yield highly accurate and reliable results at the molecular level. The test uses fluorescent probes to "paint" the HER-2 genes in the tumor cell nucleus, to see if the number of gene copies is normal or not. In addition, because this test measures genetic material, which is very stable, tissue preparation has very little effect on test outcome.

 

Ordering PathVysion From Your Lab

HER-2 status assessment is now a standard of care in the selection of therapy for invasive breast cancer.

 

Some laboratories continue to use IHC to assess HER-2 status.  To ensure your patient’s HER-2 status is determined by a FISH-based method, ask for PathVysion , specifically, on the laboratory requisition form.

 
 
 
 
Lab Professionals

Welcome

Advances in breast cancer testing and treatment have greatly improved patient outcomes. Knowing a woman's HER-2 status is an essential component to understanding how to treat breast cancer and potentially extend survival. 

 

labhome-pic.jpg

 

This web site will provide you with an overview of the HER-2 testing options currently available and why studies have consistently shown that the PathVysion HER-2 test, which employs FISH technology, can help overcome technical and interpretative limitations of other testing methods to accurately and reproducibly assess a woman's tumor for HER-2 status.

 

This section provides the information you need to know:

The Expanding Role of HER-2 Testing

HER-2 status is a strong, independent predictor of prognosis and disease free progression. Accurate assessment of HER-2 status is important in selecting appropriate therapy, including Herceptin® therapy.

 

Herceptin® (trastuzumab) is now approved in the adjuvant setting to treat early-stage HER-2 positive breast cancer, and is approved as both a first-line therapy in combination with chemotherapy and as a single agent in second- and third-line therapy.

 

The risks of incorrect HER-2 status assessment are significant: false positive assessments may lead to inappropriate medical decisions, causing a patient to undergo unnecessary therapy. Conversely, a false negative assessment may deprive a patient of potentially beneficial therapy.

 

HER-2 Testing Options

An accurate HER-2 assessment can enable the most appropriate therapy decision.

 

Tests for HER-2 status measure either gene copy number or presence of protein receptors. The two most widely used technologies worldwide and approved by the FDA in the United States are Immunohistochemistry (IHC) and Fluorescence in situ Hybridization (FISH). Chromogenic in situ Hybridization (CISH) and Silver in situ Hybridization (SISH) are two other technologies that measure HER-2 gene copy number. All of these techniques are routinely utilized on formalin-fixed, paraffin-embedded breast cancer tissue.

 

ImmunoHistoChemistry (IHC)

IHC is a technique that utilizes specific antibodies for staining protein in tissues on microscopic glass slides. Immunohistochemical staining is widely used in the diagnosis and treatement of many types of cancer.

 

The IHC test for HER-2 is a protein-based test that is used to determine the total amount of HER-2 protein receptors on the surface of the cell. The surface of the cell is "stained" with an antibody. IHC is interpreted by the intensity and percentage of cells which exhibit a brown cell surface staining. These specimens are interpreted on a numeric scale of 0 (negative for HER-2 overexpression), 1+ (negative for HER-2 overexpression), 2+ (positive for HER-2 overexpression), and 3+ (positive for HER-2 overexpression).

 

Issues regarding the accuracy of this method include:

  • The protein may be damaged by the processes of fixing and storing the tissue with  formalin and parrafin, causing variability in test outcome;
  • An assortment of antibodies are used for the testing, including laboratory developed antibodies, which lead to variability; and
  • Interpretation is relatively subjective and requires a great level of skill and expertise.

 

ihc-sampleimages.gif

Fluorescence in situ Hybridization (FISH)

 

FISH is a cytogenetic technique that can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes. It uses fluorescent probes that can be visualized with a fluorescent microscope.

 

FISH Process

fishprocess5.jpg

FISH for HER-2 is a gene-based test that measures the number of HER-2 genes in a cell. The FISH test "highlights" the HER-2 genes inside the cell, making them appear as fluorescent signals (dots) so they may be accurately counted. PathVysion® FISH also measures the number of copies of chromosome 17 in the cell. Since the HER-2 gene resides on chromosome 17, this adds several measures of control to the test. After counting the HER-2 and Chromosome 17 signals in 20 nuclei, the ratio of HER-2 to Chromosome 17 is calculated. If this value is 2 or greater, the patient is considered amplified for HER-2 or positive. If the test shows a normal gene count, she is considered HER-2 negative. FISH allows the viewer to literally count the genes, which can help overcome technical and interpretative limitations of other testing methods. In addition, because this test measures genetic material, which is very stable, tissue preparation has very little effect on test outcome. 

 

her2-pathimage.gif

Images are examples of PathVysion results. Individual results may vary.

 

Why PathVysion?

 

The PathVysion HER-2 DNA Probe Kit can be a valuable tool in the fight against breast cancer. It is a precise test that provides clinicians and their patients with highly accurate and reliable HER-2 results.

 

The PathVysion HER-2 DNA probe kit is one of the first examples of what is recognized as genomic disease management, or personalized medicine. This means that the test enables the accurate assessment of a patient's HER-2 status at the DNA level with a high degree of accuracy and helps guide doctors to make the most appropriate therapy decisions based on the patient's own genetic profile.

 

PathVysion is approved by the U.S Food and Drug Administration (FDA) for use as an aid in determining prognostic factors in patients with stage II, node-positive breast cancer, as an aid to predict disease-free and overall survival in patients with stage II, node-positive breast cancer treated with adriamycin-based chemotherapy, and assisting in Herceptin® (Trastuzumab) monoclonal antibody therapy selection.

 

PathVysion is the best test for assessing HER-2 status:

  • FISH is considered the gold standard for HER-2 testing.
  • Studies indicate PathVysion can help overcome technical and interpretative limitations of other testing methods, including IHC.
  • PathVysion exhibits better concordance than IHC between local and central laboratories
  • The Chromosome 17 probe acts as an internal control and corrects for polysomy (identifies gene amplification versus chromosome 17 polysomy) and nuclear truncation
  • FDA approved automated result scanning is available for PathVysion
  • Abbott Molecular is the clear leader in FISH technology in total US sales and research and development investment

PathVysion direct labeled FISH probes yield results at the molecular level. The test uses fluorescent probes to "paint" the HER-2 genes in the tumor cell nucleus, to see if the number of gene copies is normal or not. Interpretation is objective - results are obtained by counting the fluorescent signals (dots). In addition, because this test measures genetic material, which is very stable, tissue preparation has very little effect on test outcome.

 
 
 
 
FAQs for Patients

What is HER-2/neu?

HER-2/neu is also referred to as HER-2 or ERBB2. HER-2 stands for Human Epidermal Growth Factor Receptor 2. The HER-2 gene is one of thousands of genes that comprise every living person's genetic code and is present in the nucleus of all human cells, including breast tissue cells. The HER-2 gene provides the blueprint for the manufacture of the HER-2 protein. This protein, which is attached to the cell membrane, regulates normal cell growth and function.

 

How common is HER-2 positive breast cancer?

Abnormally high quantities of the HER-2 gene have been associated with rapid tumor cell growth, resistance to therapy, and shorter disease-free periods and overall survival. Approximately 20-25%, or 1 out of every 4, breast cancers have an abnormally high quantity of the HER-2 gene, and are considered HER-2 positive.

 

Do I need to know my HER-2 status?

Your HER-2 status can be an important factor as you and your doctor decide on your best treatment options. The NCCN (National Comprehensive Cancer Network) and ASCO (The American Society of Clinical Oncologists) recommend HER-2 assessment for all invasive breast cancers.

 

While it is recommended at initial diagnosis, HER-2 status can be determined at anytime after initial diagnosis as well. There is discussion within the medical community about whether HER-2 status should be assessed with each occurrence or recurrence of invasive breast cancer.

 

How will knowing my HER-2 status affect my therapy decisions?

Determination of HER-2 status has become an important tool in the routine assessment and management of patients with breast cancer. Abnormally high quantities of the HER-2 gene have been shown to result in more aggressive tumor behavior. HER-2 positive patients may react differently to chemotherapy. Studies have shown HER-2 positive tumors obtain greater benefit from the addition of anthracyclines and from more dose-intensive anthracycline administration than HER-2 negative tumors.

 

HER-2 amplified tumors may benefit from targeted monoclonal antibody therapies, such as Herceptin®(www.herceptin.com). 

 

How will my HER-2 status be identified?

The two most widely used technologies are ImmunoHistochemistry (IHC) and Fluorescence in situ Hybridization (FISH). The PathVysion test employs FISH technology to determine HER-2 status.

 

Does the type of HER-2 test make a difference?

Yes. An accurate HER-2 assessment guides doctors to make the most appropriate therapy decisions based on the patient's own genetic profile.

 

The IHC test is a protein-based test that is used to determine the total amount of HER-2 protein receptors on the surface of the cell. To do this, the surface of the cell is "stained" with an antibody. The surface protein measured by IHC can be damaged during preparation of the tissue sample and this may cause variability in outcome. A pathologist must judge the degree of color change in the cell to determine a HER-2 protein measurement level of 0, 1+, 2+, or 3+. Because this test is relatively subjective, the results often are confirmed by performing a FISH test.

 

PathVysion, the FISH HER-2 Test, can help overcome many of the inherent technical and interpretative limitations of other techniques, such as immunohistochemistry. The PathVysion® test yields highly accurate and reliable results at the molecular level. PathVysion "highlights" the HER-2 genes inside the cell, making them appear fluorescent so they may be accurately counted. If a patient has an abnormal number of HER-2 genes, she is considered HER-2 positive. If the test shows a normal gene count, the test results are considered HER-2 negative. PathVysion allows the physician or pathologist to literally count the genes, which can help overcome technical and interpretative limitations helping to positively identify women who are candidates for HER-2-targeted therapies.

 

What if I have already had a biopsy, but don't know if I've been screened for HER-2?

Your HER-2 results can be obtained from your biopsy tissue, at any time, by using the PathVysion test. Because the test measures the HER-2 gene at the stable DNA molecular level, results can be obtained from stored tissue samples. Ask your doctor for more information. Knowing your HER-2 status could be important for your treatment decisions.

 

What are the limitations associated with PathVysion I should be aware of?

  1. The PathVysion Kit has been optimized only for identifying and quantifying chromosome 17 and the HER-2/neu gene in interphase nuclei from formalin-fixed, paraffin-embedded human breast tissue specimens. Other types of specimens or fixatives should not be used.
  2. The performance of the PathVysion Kit was validated using the procedures provided in this package insert only. Modifications to these procedures may alter the performance of the assay.
  3. Performance characteristics of the PathVysion Kit have been established only for node positive patients receiving the designated regimens of CAF and for metastatic breast cancer patients being considered for HERCEPTIN therapy. Performance with other treatment regimens has not been established.
  4. The clinical interpretation of any test results should be evaluated within the context of the patient’s medical history and other diagnostic laboratory test results.
  5. FISH assay results may not be informative if the specimen quality and/or specimen slide preparation is inadequate.
  6. Technologists performing the FISH signal enumeration must be capable of visually distinguishing between the orange and green signals.

What are the warnings associated with PathVysion I should be aware of?

  1. For In Vitro Diagnostic Use.
  2. The PathVysion Kit is intended for use only on formalin-fixed, paraffin-embedded breast cancer tissue; it is not intended for use on fresh or non-breast cancer tissue.
  3. All biological specimens should be treated as if capable of transmitting infectious agents. The ProbeChek control slides are manufactured from human cell lines that have been fixed in 10% formalin. Because it is often impossible to know which might be infectious, all human specimens and control slides should be treated with universal precautions. Guidelines for specimen handling are available from the U.S. Centers for Disease Control and Prevention.16
  4. Exposure of the specimens to acids, strong bases or extreme heat, should be avoided. Such conditions are known to damage DNA and may result in FISH assay failure.
  5. Failure to follow all procedures for slide denaturation, hybridization and detection may cause unacceptable or erroneous results.
  6. To identify target areas, H & E staining should be conducted on every 10th slide of the same tissue block.
  7. Hybridization conditions may be adversely affected by the use of reagents other than those provided by Abbott Molecular Inc.
  8. Proper storage of kit components is essential to ensure the labeled shelf life. Assay results may be adversely affected by kit components stored under other conditions.
  9. If stored at low temperatures, 20X SSC may crystallize. If the crystals cannot be redissolved at room temperature, the solution should be discarded.
  10. If any other working reagents precipitate or become cloudy, they should be discarded and fresh solutions prepared.
  11. The DAPI Counterstain contains DAPI (4,6-diamidino-2-phenylindole) and 1,4-phenylenediamine.

    •  DAPI is a possible mutagen based on positive genotoxic effects. Avoid inhalation, ingestion or contact with skin.

    •  DAPI is a possible mutagen based on positive genotoxic effects. Avoid inhalation, ingestion or contact with skin.
  12. Fluorophores are readily photo bleached by exposure to light. To limit this degradation, handle all solutions containing fluorophores in reduced light. This includes all steps involved in handling the hybridized slide. Carry out all steps, which do not require light for manipulation (incubation periods, washes, etc.) in the dark.
  13. LSI HER-2/neu & CEP 17 DNA probe mixture contains formamide, a teratogen. Avoid contact with skin and mucous membranes.
  14. Calibrated thermometers are required for measuring temperatures of solutions, water baths and incubators.
  15. Always verify the temperature of the pretreatment solution, denaturation solution and wash buffers prior to each use by measuring the temperature of the solution in the coplin jar with a calibrated thermometer.
  16. All hazardous materials should be disposed of according to your institution's guidelines for hazardous disposal. The following are European Community (EC) risk and safety information. Restricted to professional users.
 
 
 
 
faqs for Clinicians & Laboratory Professionals

What is HER-2/neu? 

HER-2/neu, also known as ERBB2, is a gene that plays a key role in the regulation of cell growth. The HER-2 gene is amplified in human breast, ovarian, and other cancers, and plays an important role in the progression of many tumors. This amplification is characterized by increased transcription of mRNA and subsequent overexpression of the gene product. Approximately 20-25% of breast cancers have amplification of the HER-2 gene. Detection of HER-2 amplification is important in the diagnosis, prognosis, selection of appropriate therapy and prediction of therapeutic outcome in certain cancers.

 

How important is HER-2 amplification in the treatment of breast cancer? 

HER-2 amplification and subsequent overexpression of the gene product have emerged as a key prognostic and therapeutic marker in breast cancer. Amplification of this gene is found to be associated with rapid proliferation, shorter disease-free survival and poorer overall survival in both node-negative and node-positive ductal breast cancers. With the advent of monoclonal targeted therapies such as Herceptin® ® (Trastuzumab), accurate HER-2 assessment is a critical component in determining appropriate therapy selection. Both the National Comprehensive Cancer Network (NCCN®) and the American Society of Clinical Oncology (ASCO®) recommend HER-2 assessment for all invasive breast cancers. 

 

What is the connection between HER-2 status and the selection of adriamycin-based therapy?

An interaction between HER-2 amplification and response to adriamycin-based therapies has been demonstrated. Specifically, HER-2 amplification is associated with improved response to intensified adriamycin-based chemotherapies. Clinical trials demonstrate that patients with HER-2 amplification may benefit from dose intensive adriamycin-based therapy based on disease-free and overall survival. Dose intensification of adriamycin did not benefit women without HER-2 amplification.

 

What is the connection between HER-2 status and Herceptin® therapy selection? 

The Herceptin package insert states that detection of HER-2 status is necessary for selection of patients appropriate for Herceptin therapy. In a single-arm study of Herceptin as a single agent no relapsed metastatic breast cancer, the overall response rate in 173 patients with FISH positive tumors tested was 19%, whereas no objective responses were observed in patients with FISH( -) tumors. All patients' specimens had previously demonstrated over expression of HER-2 at the 2+ or 3+ level by IHC. For more information on HER-2 testing and Herceptin, visit www.herceptin.com.

 

Why is accurate assessment of HER-2 status essential?

HER-2 status is a strong, independent predictor of prognosis and disease free progression. Accurate assessment of HER-2 status is important in selecting appropriate therapy, including Herceptin therapy.

 

The risks of incorrect HER-2 status assessment are significant: false positive assessments may lead to inappropriate medical decisions, causing a patient to undergo unnecessary therapy. Conversely, a false negative assessment may deprive a patient of potentially beneficial therapy. 

 

Should one be assessing HER-2 at the DNA level or the gene product (protein) level?

Extra copies of the HER-2 gene drive excess protein on the surface of the cell. HER-2 assessment at a direct molecular level by FISH has been shown to be more accurate than IHC antibody-based tests. Responses in IHC 2+ and 3+ MBC patients treated with single-agent Herceptin were seen exclusively in FISH positive patients.

 

What are the most accurate methods available for HER-2 status assessment?

Tests for HER-2 status measure either gene copy number or presence of protein receptors. The two most widely used technologies and the only two approved by the FDA in the United States are Immunohistochemistry (IHC) and Fluorescence in situ Hybridization (FISH). Chromogenic in situ Hybridization (CISH) and Silver in situ Hybridization (SISH) are two other technologies that measure HER-2 gene copy number. All of these techniques are routinely utilized on formalin-fixed, paraffin-embedded breast tissue.

 

While IHC has been a widely used technique, questions have been raised as to which technique, either FISH or IHC, yields the most reliable and accurate detection of HER-2 status. From the clinical pathology perspective, IHC inherently possesses several key shortcomings that can interfere with the accurate assessment of HER-2 status. Formalin-fixation of tissues is important in stabilizing cellular morphology and it introduces variables for IHC determination that are not within the laboratory's power to control. Specifically, formalin-fixation can cause destruction of the HER-2 epitope, which can lead to false negative IHC results. To help minimize this shortcoming, antigen retrieval techniques have now been employed. However, these antigen retrieval techniques cause a significant number of false positive IHC results.

 

Direct FISH technology targets stable DNA at a molecular level within formalin-fixed, paraffin-embedded specimens thereby overcoming the misclassifications associated with IHC. Direct FISH is a very sensitive method that yields higly accurate, reliable and reproducible results simply from counting the number of fluorescent signals. From these unambiguous results, the pathologist can make accurate HER-2 assessments, eventually leading oncologists and patients to the most efficacious selection of therapy.

 

What is the PathVysion HER-2 DNA Probe Kit?

The PathVysion HER-2 DNA Probe Kit utilizes patented, direct labeled Fluorescent in situ Hybridization DNA probes to yield definitive results at the molecular level. FISH is a molecular genetic technique employing a fluorescent DNA probe that produces a bright microscopic signal when it selectively attaches to the gene or chromosome specific complementary DNA. The PathVysion HER-2 DNA Probe Kit contains two probes. One probe is directed to the HER-2 gene and the second DNA probe attaches itself to the chromosome 17 centromere. The signals appear as orange and green fluorescent dots for the HER-2 gene and chromosome 17, respectively. This enables a ratio of HER-2 to chromosome 17 signals to be calculated thereby correcting for aneuploidy of chromosome 17, which may not represent an amplified HER-2 state. Through this direct labeled FISH technology, the PathVysion HER-2 assay provides an accurate, reliable and reproducible means for assessing HER-2 status in breast cancer.

 

What is direct labeled FISH? 

FISH has become a main-stream technology routinely used in diagnostic laboratories. Two kinds of FISH technology are commercially available - - direct labeled and indirect labeled.

 

Direct labeled FISH uses probes that have been pre-labeled with a specific fluorophore, allowing the fluorescent signal to be bound to the target in a single hybridization step. This method is more advanced, has fewer steps, is easier-to-read and has faster time-to-result. With PathVysion, HER-2 assessment is accomplished with a sequence specific DNA probe for the HER-2 gene and a second, separate DNA probe labeled in a different fluorophore for the repetitive sequences at the centromere of chromosome 17 (the HER-2 gene resides on chromosome 17). Direct FISH is Abbott Molecular's specialty.

 

Indirect labeled FISH involves probes prelabeled with a hapten, most commonly digoxigenin. After hybridization, fluorochrome-labeled antibodies to the hapten are used for probe detection. As an example, anti-digoxigenin, provides a "sandwiching" assay. Indirect FISH also requires additional blocking reagents, amplification steps, and has been reported to have a higher degree of background fluorescence problems. The need for extra handling post-hybridization can cause disruptions in work-flow within a busy clinical laboratory; therefore, direct label FISH represents a superior technology and offers many advantages.

 

Why does the PathVysion HER-2 DNA kit include a probe for chromosome 17?

The HER-2 gene is located on chromosome 17. The cut-off between normal and abnormal is set by biology; more than one copy of the HER-2 gene per chromosome 17 is abnormal. Determination of the ratio of HER-2 gene copy number to chromosome 17 copy number is useful in the discrimination of aneusomy of chromosome 17 from true HER-2 gene amplification. In the case of aneusomy, more than the normal number of two copies of chromosome 17 are present in the cell and consequently, more than the normal number of two copies of the HER-2 gene are present. This result does not represent HER-2 amplification. Therefore a ratio of HER-2 to chromosome 17 is used in the cutoff of the PathVysion assay.

 

The chromosome 17 control probe also serves as a methodologic internal control for hybridization and controls for "nuclear truncation". When dealing with solid tumor tissue, nuclear truncation may occur. Most nuclei are roughly 10 microns thick, with a 4-6 micron section taken to be tested. The chromosome 17 probe is utilized to assess truncation and ensure the appropriate nuclei are evaluated in assessing HER-2 amplification.

 

What clinical trials have been conducted on the PathVysion HER-2 assay?

The pivotal clinical trial demonstrated that HER-2 amplification as determined by the PathVysion HER-2 DNA Probe could be used to identify those patients with a poor prognosis and furthermore, more likely to benefit from dose intensification of adriamycin-based therapies. The PathVysion HER-2 assay reliably detected HER-2 amplification, demonstrating a significant interaction between CAF (cyclophosphamide, doxorubicin, 5-fluorouracil) and amplified HER-23.

 

Herceptin package insert data from a retrospective analysis of 660 patients enrolled in the Herceptin clinical studies (all scoring 2+ or 3+ by the Clinical Trial IHC assay) demonstrate that the clinical benefits of Herceptin were greater in patients whose tumors tested FISH(+) than those that were FISH( - ). In the singlearm study of Herceptin as a single agent, the overall response rate in patients whose tumors tested as FISH(+) was 20%, while there were no responses in those who tested as FISH( - ).

 

Why can it be beneficial to specifically request the PathVysion HER-2 DNA direct label FISH when assessing HER-2 status?

HER-2 status assessment is now considered the standard of care in the selection of therapy for invasive breast cancer, and some laboratories continue to use IHC to assess HER-2 status. PathVysion is the only method of HER-2 assessment that is FDA approved for use as an aid in determining prognostic factors in patients with stage II, node-positive breast cancer, as an aid to predict disease-free and overall survival in patients with stage II, node-positive breast cancer treated with adriamycin-based chemotherapy, and assisting in Herceptin (Trastuzumab) monoclonal antibody therapy selection.

 

Can the PathVysion HER-2 assay be performed in the average lab?

Yes. Most major reference labs, cancer centers and many local hospitals now perform the PathVysion HER-2 assay as their primary method of assessing HER-2 status. The protocol for the PathVysion HER-2 assay can be easily performed at any laboratory that currently performs IHC testing.

 

How do I explain the benefits of the PathVysion HER-2 assay to my patients?

Results of the PathVysion HER-2 asssay can be best explained to patients in terms of how they directly affect the therapy they are most likely to receive. When a breast cancer patient knows her HER-2 status, the appropriate therapy choices become clear.

 

Which patients should receive the PathVysion HER-2 assay?

Testing for HER-2 plays a key role in the management of invasive breast cancer. ASCO Guidelines and NCCN Practice guidelines recommend HER-2 testing for all patients with invasive breast cancer.  NCCN Practice guidelines indicate that patients should be tested by either IHC or FISH, but that all IHC 2+ results should be confirmed by FISH.

 

Where can I obtain more information about the PathVysion HER-2 assay?

More information is currently available on PathVysion HER-2 from Abbott Molecular, Inc., including technical information, clinical trial data (package insert), a Patient Q&A brochure, a laboratory brochure. For more information, please contact Abbott Molecular today at 1-800-553-7042.

 
 
 
 
Links

 

Additional Breast Cancer and Patient Resources:

 

American Society of Clinical Oncology

The American Society of Clinical Oncology (ASCO) is a non-profit organization, with overarching goals of improving cancer care and prevention and ensuring that all patients with cancer receive care of the highest quality.

 

BreastCancer.org

Breastcancer.org is a nonprofit organization dedicated to providing the most reliable, complete, and up-to-date information about breast cancer. Their mission is to help women and their loved ones make sense of the complex medical and personal information about breast cancer, so they can make the best decisions for their lives.

 

Breast Treatment Task Force

Breast Treatment Task Force is a groundbreaking program providing free surgeries and treatments to patients without access to health insurance.

 

Cancer Care

CancerCare is a national nonprofit organization whose mission is to provide free, professional support services to anyone affected by cancer: people with cancer, caregivers, children, loved ones, and the bereaved. CancerCare programs — including counseling, education, financial assistance and practical help — are provided by trained oncology social workers and are completely free of charge.

 

College of American Pathologists

The College of American Pathologists, the leading organization of board-certified pathologists, serves patients, pathologists, and the public by fostering and advocating excellence in the practice of pathology and laboratory medicine.

 

Dr. Susan Love Research Foundation

The Mission of the Dr. Susan Love Research Foundation is to eradicate breast cancer and improve the quality of women's health through innovative research, education and advocacy.

 

Living Beyond Breast Cancer

Living Beyond Breast Cancer is dedicated to assisting breast cancer patients, whether newly diagnosed, in treatment, recently completed treatment, are years beyond or are living with advanced (metastatic) disease.

 

National Breast Cancer Coalition

The National Breast Cancer Coalition is the nation's largest breast cancer advocacy group with hundreds of member organizations and tens of thousands of individual members. 

 

NBCC's sister organization, the National Breast Cancer Coalition Fund, empowers and trains members to be effective in every aspect of the fight to end breast cancer. It gives women the tools to make their own informed decisions and to work beside legislative, scientific and clinical decisionmakers.

 

National Coalition for Cancer Survivorship

The National Coalition for Cancer Survivorship is the oldest survivor-led cancer advocacy organization in the country and a highly respected authentic voice at the federal level, advocating for quality cancer care for all Americans and empowering cancer survivors.

 

National Comprehensive Cancer Network

The National Comprehensive Cancer Network (NCCN), a not-for-profit alliance of 21 of the world's leading cancer centers, is dedicated to improving the quality and effectiveness of care provided to patients with cancer. The primary goal of all NCCN initiatives is to improve the quality, effectiveness, and efficiency of oncology practice so patients can live better lives.

 

Oncology Nursing Society

The Oncology Nursing Society (ONS) is a professional organization of more than 33,000 registered nurses and other healthcare providers dedicated to excellence in patient care, education, research, and administration in oncology nursing.

 

The Breast Cancer Research Foundation

The Breast Cancer Research Foundation is a not-for-profit organization whose mission is to achieve prevention and a cure for breast cancer by providing critical funding for innovative clinical and genetic research at leading medical centers worldwide, and increasing public awareness about good breast health.

 

Vital Options International

Vital Options® International TeleSupport® Cancer Network is a not-for-profit cancer communications, support and advocacy organization whose mission is to facilitate a global cancer dialogue by using communications technology to reach every person touched by cancer.

 

The Wellness Community

The Wellness Community is an international non-profit organization dedicated to providing education and resources to people affected by cancer—helping them learn vital skills that will enable them to regain control, reduce isolation and restore hope regardless of the stage of their disease.

 

Young Survival Coalition

The Young Survival Coalition (YSC) is an international, non-profit network of breast cancer survivors and supporters dedicated to the concerns and issues that are unique to young women and breast cancer. Through action, advocacy and awareness, the YSC seeks to educate the medical, research, breast cancer and legislative communities and to persuade them to address breast cancer in women aged 40 and under.

 
 
 
 
GLOSSARY

 

Adjuvant chemotherapy

Anticancer drugs used in combination with surgery and/or radiation as an initial treatment before there is detectable spread of the cancer, to prevent or delay recurrence.

 

Centromere

The constricted region usually near the center of a chromosome (at the joining point of chromosome arms) consisting of highly repeated DNA sequences.

 

Chromosome

Structure in a cell on which genes are located, consisting of a highly compacted stretch of DNA.

 

Chromosome Abnormalities

These can be either structural or numerical. Structural abnormalities include translocations, deletions or insertions, duplications and amplifications. Numerical abnormalities are either a gain or loss of chromosomes.

 

DNA

An acronym for deoxyribonucleic acid. DNA contains all the information necessary for any organism to develop and function. The four chemical building blocks of DNA are Adenine, Thymine, Cytosine and Guanine.

 

DNA Probe

A synthetic segment of DNA specifically designed to be complementary to a target sequence of chromosomes, genes or gene sequences. For FISH, the synthetic piece of DNA is coupled with a fluorescent indicator, or tag, so that the chromosomes or genes it binds (or hybridizes) to can be directly visualized using a fluorescent microscope.

 

Fluorescence in situ Hybridization (FISH)

A technique utilizing a fluorescent-labeled DNA probe to detect a particular chromosome or gene.

 

Fluorescence

Fluorescent signals in FISH technology are visualized using a fluorescent microscope. Fluorescence is luminescence caused by the absorption of radiation at one wavelength followed by nearly immediate reradiation usually at a different wavelength and that ceases almost at once when the incident radiation stops.

 

Gene

A segment of nucleic acid that contains the information necessary to produce a functional product, usually a protein. Genes are the biologic units of heredity. They interact with each other to influence physical development and behavior.

 

Gene amplification

Increase in the normal copy number of a gene.

 

Hybridization (e.g., DNA hybridization)

The process of joining two complementary strands of DNA. 

in situIn the natural or original position

 

Metastatic

Metastatic breast cancer refers to cancer that has left the breast and spread to distant sites.

 

Nucleic Acid

A nucleic acid is a complex, high-molecular weight molecule composed of nucleotide chains that convey genetic information. The most common nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Nucleic acids are found in all living cells and viruses.

 
 
 
 

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