What is it? The INFORM HER2 Dual ISH DNA Probe Cocktail is a laboratory test that uses DNA probes, with color-forming (chromogenic) dyes attached, to count how many copies of the HER2 gene, located on chromosome 17, are present in a tissue sample. This test, which uses a variation of fluorescence in situ hybridization (ISH), is based on color detection, rather than fluorescence, and is used with samples of tumor tissue removed from breast cancer patients. This test is used by a doctor (pathologist) trained to identify diseases by studying slices of cells and tissues placed on microscope slides. If the test result indicates that too many copies of the HER2 gene (gene amplification) are present in the cancer cell, then a breast cancer patient may be eligible for treatment with the anti-cancer drug Herceptin® (Trastuzumab).

1. INFORM HER2 Dual ISH DNA Probe Cocktail assay
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2. INFORM HER2 Dual ISH DNA Probe Cocktail assay
INFORM HER2 Dual ISH is a fully automated assay
for determining HER2 gene status in breast and
gastric cancer tissue specimens
Fully Automated
• Provides a rapid time to result and reduces resource requirements
– ~12 hour run time
– Baking Through Staining
• Code, Load and Go
Easily Interpreted Using Brightfield Microscopy
• Allows for review of the entire tissue specimen and not just a selected area
– See the HER2 gene status in the context of the tumour morphology
– Ensure scoring of invasive carcinoma versus ductal carcinoma in situ
– Identify tumour heterogeneity
• Produces an archivable result
– Staining results do not fade over time
Robust and Reproducible
• Flexible protocol to address variations in preanalytical conditions
• Highly concordant with FISH
• Highly reproducible between laboratories and pathologists HER2 (black) and Chr17 (red) 60X
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3. INFORM HER2 Dual ISH DNA Probe Cocktail assay
Overview of the INFORM HER2 Dual ISH assay
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cohybridisation
In the INFORM HER2 Dual ISH DNA Probe Cocktail assay the cocktail (A) of the DNP labeled HER2
probe and DIG labeled Chr17 probe are cohybridised on the tissue sample. The HER2 gene is detected
by a dinitrophenyl (DNP) labeled probe (A) and visualised utilising ultraView Silver ISH DNP (SISH)
detection (B). The Chromosome 17 centromere is targeted with a digoxigenin (DIG) labeled probe (A)
and detected using ultraView Red ISH DIG detection (C). Dual ISH staining utilising this Dual Detection
results in visualisation via light microscopy in which HER2 appears as discrete black signals (SISH) and
Chr17 as red signals (Red ISH).
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4. INFORM HER2 Dual ISH DNA Probe Cocktail assay
INFORM HER2 DUAL ISH DNA Probe Cocktail
Intended Use
Ventana Medical Systems (Ventana) INFORM HER2 Dual ISH DNA Probe Cocktail is
designed to quantitatively detect amplification of the HER2 gene via two-color
chromogenic ISH in formalin-fixed, paraffin-embedded human breast and gastric
cancer, including the gastroesophageal junction, tissue specimens by light
microscopy
The INFORM HER2 Dual ISH DNA Probe Cocktail is indicated as an aid in the
assessment of patients for whom Herceptin® (trastuzumab) treatment is being
considered
This product should be interpreted by a qualified reader in conjunction with
histological examination, relevant clinical information, and proper controls
This reagent is intended for in vitro diagnostic (IVD) use
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5. INFORM HER2 Dual ISH DNA Probe Cocktail assay
The INFORM HER2 Dual ISH is fully automated on
all BenchMark series instruments
Provides a rapid time to result and
reduces resource requirements
• ~12 hour run time
Baking Through Staining
• Code, Load and Go
On all BenchMark series instruments
• BenchMark GX system
• BenchMark XT system
• BenchMark ULTRA system
BenchMark XT BenchMark ULTRA
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6. INFORM HER2 Dual ISH DNA Probe Cocktail assay
The INFORM HER2 Dual ISH is easily interpreted
using brightfield microscopy
Breast carcinoma with low level HER2 amplification
Allows for review of the entire tissue
specimen, not just a selected area
– Use 20X, 40X or 60X objectives
rather than 100X oil
– See the HER2 gene status in the
context of the tumour morphology
– Ensure scoring of invasive carcinoma
versus ductal carcinoma in situ
– Identify tumour heterogeneity
Produces an archivable result
HER2 (black) and Chr17 (red) 60X – Staining results do not fade over
time
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7. INFORM HER2 Dual ISH DNA Probe Cocktail assay
INFORM HER2 Dual ISH has a flexible protocol to
address variations in preanalytical conditions
• Recommended slide preparation
– 10% neutral buffered formalin (NBF) for 6 to 48 hours
– Embedded in paraffin
– Sectioned at approximately 4 microns
• To compensate for tissue variations, the assay has been designed with user
selectable protocol steps to allow for optimization
• <6% of slides may need to be restained
• Recent studies suggest the majority of inconclusive HER2 gene assay
results relate to pre-analytical factors
– Under fixation
– Over fixation
– Delay to fixation
– Fixative
– Tissue thickness
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8. INFORM HER2 Dual ISH DNA Probe Cocktail assay
HER2 Dual ISH is highly concordant to
Vysis PathVysion HER2 FISH on Breast
• HER2 Dual ISH vs Vysis PathVysion HER-2 DNA Probe Kit FISH in
breast tissue
• First pass staining success rate
– FISH 60% (80% after second attempt)
– Dual ISH >93%
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9. INFORM HER2 Dual ISH DNA Probe Cocktail assay
HER2 Dual ISH is highly concordant to
Dako HER2 FISH PharmDx on Gastric
• HER2 Dual ISH vs Dako HER2 FISH PharmDx kit in gastric tissue
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10. INFORM HER2 Dual ISH DNA Probe Cocktail assay
INFORM HER2 Dual ISH can be combined with
the Ventana fully automated IHC breast panel
Ventana breast panel antibodies
are all Rabbit Monoclonal
• PATHWAY HER2 (4B5)
• CONFIRM ER (SP1)
• CONFIRM PR (1E2)
•CONFIRM Ki-67 (30-9)
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11. INFORM HER2 Dual ISH DNA Probe Cocktail assay
Ventana the only provider of fully automated
HER2 ISH and IHC solutions
Patient Breast or Gastric Tissue Sample
Pathway 4B5 INFORM HER2 DISH
0 1+ 2+ 3+ <2 ≥2
Brightfield
Negative Negative Reflex Positive Negative Positive
INFORM HER2 DISH
Report as HER2 positive to
Oncologist for Herceptin
<2 ≥2 Consideration
Negative Positive
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12. INFORM HER2 Dual ISH DNA Probe Cocktail assay
INFORM HER2 Dual ISH is a fully automated assay
for determining HER2 gene status in breast and
gastric cancer tissue specimens
Fully Automated
• Provides a rapid time to result and reduces resource requirements
– ~12 hour run time
– Baking Through Staining
• Code, Load and Go
Easily Interpreted Using Brightfield Microscopy
• Allows for review of the entire tissue specimen and not just a selected area
– See the HER2 gene status in the context of the tumour morphology
– Ensure scoring of invasive carcinoma versus ductal carcinoma in situ
– Identify tumour heterogeneity
• Produces an archivable result
– Staining results do not fade over time
Robust and Reproducible
• Flexible protocol to address variations in preanalytical conditions
• Highly concordant with FISH
• Highly reproducible between laboratories and pathologists HER2 (black) and Chr17 (red) 60X
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