Leica Biosystems Bond Oracle HER2 IHC System - Manual (Page 10)

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Leica Biosystems Bond Oracle HER2 IHC System Instructions for Use TA9145 EN-CE-Rev_E 18/06/2013
HER2 Control Slide – HER2 Primary Antibody
Each of the supplied HER2 Control Slides contains four formalin-fixed, paraffin-embedded
human breast cancer cell line cores with staining intensity scores of 0, 1+, 2+ and 3+. One slide
must be included in each test run (ie slide tray). The correct evaluation of the HER2 Control
Slide supplied by Leica Biosystems indicates the validity of the test (refer to Bond Oracle HER2
IHC System Interpretation Guide). The HER2 Control Slides supplied with this system validate
reagent performance only and do not verify tissue preparation.
In-house Positive Control Tissue – HER2 Primary Antibody
If in-house positive control tissue components are used, they should be biopsy or surgical
specimens fixed, processed and embedded as soon as possible in the same manner as the
patient sample(s). Positive tissue controls are indicative of correctly prepared tissues and
valid staining techniques. At least one positive control component for each test run should be
included. The positive control section should demonstrate weak positive staining so as to define
subtle changes in primary antibody sensitivity.
Note:
Known positive control tissue components should only be utilized for monitoring the correct
performance of processed tissues together with test reagents, NOT as an aid in formulating
a specific interpretation of patient samples. If the positive control tissue fails to demonstrate
appropriate positive staining, results obtained with patient specimens should be considered
invalid.
A multi tissue control block containing tumors representing all 4 HER2 grades may also be
effectively utilized as appropriate in-house control material.
In-house Negative Control Tissue Component – HER2 Primary Antibody
If in-house negative control components are used, they should be fresh biopsy or surgical
specimens fixed, processed and embedded as soon as possible in the same manner as the
patient sample(s). Use of control tissue, known to be HER2 oncoprotein negative, with each
staining run verifies the specificity of the primary antibody and provides an indication of any
nonspecific background staining. The variety of different cell types present in most tissue
sections offers internal negative control sites (this should be verified by the user). Normal breast
ducts unassociated with tumor may provide a reference to the validity of the assay. If specific
staining occurs in the internal negative control tissue, results with the patient specimens should
be considered invalid.
The use of multi-tissue control block representing all four HER2 grades may be utilized for the
purposes of negative and postive control tissues.
Patient Tissue – HER2 Negative Control
Use the supplied HER2 Negative Control in place of the HER2 Primary Antibody on a
corresponding section for each patient test to evaluate nonspecific staining and allow accurate
interpretation of specific HER2 oncoprotein staining at the antigenic site.
Patient Tissue – HER2 Primary Antibody
Positive staining intensity should be assessed within the context of any nonspecific background
staining with the HER2 Negative Control. As with any immunohistochemical test, a negative
result means that the antigen was not detected, not that the antigen was absent in the cells/
tissue assayed. Refer to Slide Screening Order Rationale, Limitations, Performance Evaluation
and Immunoreactivity for specific information regarding Bond Oracle HER2 IHC System
immunoreactivity.
Assay Verification
Prior to the initial use of any antibody or staining system in a diagnostic procedure, the user
should verify the antibody’s specificity by testing it on a series of in-house tissues with known
immunohistochemical positive and negative profiles. Refer to Quality Control as previously outlined
and the quality control requirements of the CAP Certification Program for Immunohistochemistry