Leica Biosystems Bond Oracle HER2 IHC System - Manual (Page 13)

Page 13 of 23
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Leica Biosystems Bond Oracle HER2 IHC System Instructions for Use TA9145 EN-CE-Rev_E 18/06/2013
English
Nonspecific staining, if present, usually has a diffuse appearance. Sporadic staining of
connective tissue may also be observed in sections from excessively formalin-fixed tissues.
Use intact cells for interpretation of staining results. Necrotic or degenerated cells often stain
nonspecifically (16). False-positive results may be seen due to non-immunological binding of
proteins or substrate reaction products. They may also be caused by endogenous enzymes
such as pseudoperoxidase (erythrocytes) or endogenous peroxidase (cytochrome C),
depending on the type of immunohistochemical stain used.
Tissues from patients infected with Hepatitis B virus and containing Hepatitis B virus surface
antigen (HBsAg) may exhibit nonspecific staining with horseradish peroxidase (17).
Unexpected immunohistochemical staining, or variations in the staining, may be as a result
of alterations in the expression levels of the encoding genes or antigens. Any change in
expected staining patterns should be interpreted in association with all other diagnostic
investigations.
The interpretation of immunohistochemical staining should be complemented by morphological
studies and the use of suitable control material, and should be evaluated within the context of
the patient’s clinical history and other any diagnostic tests by a qualified pathologist.
The performance of the assay (ie assessments of adequacy of both positive and negative
controls) and the interpretation of any immunohistochemical staining or its absence must
be carried out in an appropriately accredited/licensed laboratory under the supervision of a
suitably qualified and experienced pathologist, who is responsible for the overall assessment
of the immunohistochemical assay and its interpretation.
B. Product Specific Limitations
This product is not intended for use in flow cytometry. Performance characteristics have not
been determined for flow cytometry.
False negative results may be seen as a result of the degradation of antigens in the tissue
section. Slides required for HER2 oncoprotein evaluation and tumor verification should be
prepared at the same time. To preserve antigenicity, tissue sections mounted on slides
(Leica BOND Plus Slides – product code S21.2113) should be stained within 4–6 weeks
of sectioning when held at room temperature (20–25 °C). Following sectioning, slides are
recommended to be incubated for 12–18 hours at 37 °C. Sections which require further
adherence may be incubated at 60 °C for a further hour.
Minimal natural variation of immunohistochemical profile will be seen between growth batches
of cell lines utilized within the Bond Oracle HER2 IHC System. This natural variation is well
within acceptable tolerance levels of a biological entity and does not affect the interpretation
or performance of the system.
Characterization of the cell lines using both flow cytometry and in situ hybridization
as presented in Table 5 are also subject to natural biological variation. Technical and
interpretational variation of control cell lines as assessed by fluorescent in situ hybridization
is also reported (18).
Assessment of the HER2 Control Slides should take into account all relevant expiry dates.
Store the Bond Oracle HER2 IHC System at 2–8 °C. Do not freeze. Return to 2–8 °C
immediately after use. Any deviations from these conditions will invalidate the assay.
Do not replace Bond Oracle HER2 IHC System reagents with any other components either
supplied by Leica Biosystems or by other manufacturers. To do so will invalidate the assay.
It is essential that all of the steps outlined in sections C to E (Procedure) are performed in the
prescribed order. Any deviation from this order will invalidate the assay.
It is essential that tissues fixed only in formalin-based fixatives be used in the assay. The use
of any other type of fixative will invalidate the assay.
Tissue sections cut outside of the recommended thickness range have not been validated.
The use of any other section thickness may invalidate the assay.