Leica Biosystems HER2 FISH System - 30 Test - Manual (Page 4)

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Leica Biosystems Leica HER2 FISH System - 30 Test Instructions for Use TA9217 EN-CE-Rev_D 08/04/2013
The characteristic morphology of the tissue and the spatial distribution of oncogene copies in
individual uncultured primary breast carcinomas are retained. Aberrations in chromosome 17
copy number (aneusomy) are also commonly found in breast tumors. These may present as
chromosome deletions or gains (polysomy). This chromosomal variation has critical impact on
the interpretation and reporting of HER2 gene amplification status. Therefore, measurement of
chromosome 17 copy number in conjunction with HER2 is critically important (4).
The Leica HER2 FISH System - 30 Test contains the LSI HER2 DNA probe, a 226 Kb
SpectrumOrange ™ directly labeled fluorescent DNA probe specific for the HER2 gene
locus (17q11.2-q12) and the CEP17 DNA probe, a 5.4 Kb SpectrumGreen ™ directly labeled
fluorescent DNA probe specific for the alpha satellite DNA sequence at the centromeric region
of chromosome 17 (17p11.1-q11.1). The probe solution has been specially formulated and
validated for use on the Leica BOND-MAX and BOND-III System and should not be modified
or used in a manual setting.
Clinical Concordance Summary Leica BOND-MAX System
The Leica HER2 FISH System - 30 Test was developed to provide a fully automated alternative
to current methodologies used to determine HER2 gene amplification status. The performance
of the Leica HER2 FISH System - 30 Test on the Leica BOND-MAX System was evaluated in an
independent study comparing the results of the Leica HER2 FISH System - 30 Test to the Abbott
Molecular PathVysion HER-2 DNA Probe Kit Assay on 300 breast tumor specimens. None of
these tumor specimens were obtained from patients in the Herceptin clinical studies. The results
indicated a 99.33% concordance in a 2x2 analysis (95% confidence intervals of 97.61–99.92%).
The concordance data also indicates that a positive result with the Leica HER2 FISH System -
30 Test is highly likely to correspond with a positive result on the Abbott Molecular PathVysion
HER-2 DNA Probe Kit assay. The Leica HER2 FISH System - 30 Test is interpreted as negative
for HER2 gene amplification when the HER2:CEP17 gene ratio is less than 2.0 and positive
when the HER2:CEP17 gene ratio is greater than or equal to 2.0. Equivocal (borderline) results,
where the HER2:CEP17 gene ratio is between or equal to 1.8-2.2, should be interpreted with
caution. Count an additional 20 nuclei and recalculate the ratio.
Clinical Concordance Summary Leica BOND-III System
The Leica HER2 FISH System - 30 Test was developed to provide a fully automated alternative
to current methodologies used to determine HER2 gene amplification status. The performance
of the Leica HER2 FISH System - 30 Test on the Leica BOND-III System was evaluated in an
independent study comparing the results of the Leica HER2 FISH System - 30 Test to the Abbott
Molecular PathVysion HER-2 DNA Probe Kit Assay on 300 breast tumor specimens. None of
these tumor specimens were obtained from patients in the Herceptin clinical studies. The results
indicated a 99.67% concordance in a 2x2 analysis (95% confidence intervals of 98.16–99.99%).
The concordance data also indicates that a positive result with the Leica HER2 FISH System -
30 Test is highly likely to correspond with a positive result on the Abbott Molecular PathVysion
HER-2 DNA Probe Kit assay. The Leica HER2 FISH System - 30 Test is interpreted as negative
for HER2 gene amplification when the HER2:CEP17 gene ratio is less than 2.0 and positive
when the HER2:CEP17 gene ratio is greater than or equal to 2.0. Equivocal (borderline) results,
where the HER2:CEP17 gene ratio is between or equal to 1.8-2.2, should be interpreted with
caution. Count an additional 20 nuclei and recalculate the ratio.
Principle of Procedure
The Leica HER2 FISH System - 30 Test contains components required to complete a fluorescence
in situ hybridization based staining procedure for formalin-fixed, paraffin-embedded tissues.
Following appropriate pretreatment, incubation with the ready-to-use LSI HER2/CEP17 Dual
Probe and appropriate stringency washing, tissue sections are then dehydrated and mounted
with DAPI. Results are interpreted by fluorescence microscopy using the recommended filters
at the appropriate wavelengths.