Leica TA9217 사용자 설명서

  

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Leica Biosystems  Leica HER2 FISH System - 30 Test Instructions for Use TA9217 EN-CE-Rev_D 08/04/2013

It is recommended that a Leica HER2 FISH Control Slide is included in each test run to monitor 

assay performance and to assess the accuracy of signal enumeration. Control slides should be 

run for each staining batch on the Leica BOND-MAX and BOND-III System and with each new 

reagent lot.  In addition, individual users may choose to use their own control material.
Assess control slide quality and perform signal enumeration according to the instructions in the 

Signal Assessment and Enumeration section.  The criteria for slide quality must be satisfied 

and the HER2:CEP17 ratio results should be within the established ranges for acceptable test 

performance. See Table 3 for acceptance criteria of the Leica HER2 FISH Control Slides.

Cell Line

Bond Oracle 
HER2 IHC 
System Profile

HER2 
Receptor Load 
per cell*

Leica HER2 FISH System - 30 Test  
HER2:CEP17 Acceptance Criteria

SKBr-3

3+

4.3 x 10

5

HER2 amplification is observed

MDA-MB-453

2+

1.4 x 10

5

HER2/CEP17 gene ratio should be between  
1.5 – 2.5

MDA-MB-175

1+

6.3 x 10

4

HER2 amplification is not observed

MDA-MB-231

0

9.3 x 10

3

HER2 amplification is not observed

If assay controls fail, FISH results for that case should not be reported. If control slides fail to 

meet the slide acceptance criteria, the Leica HER2 FISH System - 30 Test may have performed 

inadequately.  In  this  instance,  a  repeat  test  with  fresh  control  slides  and  patient  specimen 

slide(s) will be required. If the results are outside of the specified range, but the control slides 

meet the acceptance criteria for quality, repeat screening of the same slide may be appropriate 

since the enumeration may not have been performed correctly. Consult the troubleshooting 

guide (Table 6) in the event of hybridization failure, with either the specimen or control slide(s).
For clinical specimens, when interpretation of the hybridization signal is difficult and there is 

insufficient specimen sample for re-assay, the test is uninformative. If there are insufficient cells 

for analysis, the test is uninformative.
Patient specimens should be controlled according to standard laboratory operating procedures.  

Signal quality and enumeration results should be documented on an appropriate report form. 

FISH is a technique that requires specialized training in all aspects of the procedure (including 

the selection of appropriate reagents, tissue, fixation, processing and slide preparation) and 

interpretation.  Tissue  staining  is  dependent  on  the  handling,  fixation  and  processing  of  the 

tissue prior to staining. Improper fixation, freezing, thawing, washing, drying, heating, sectioning 

or contamination with other tissues or fluids may produce morphological artifacts, nucleic acid 

degradation, background fluorescence or false negative results. Inconsistent results may be 

due  to  variations  in  fixation,  embedding  methods,  or  inherent  irregularities  within  the  tissue 

(21). Excessive or incomplete counterstaining may also compromise correct interpretation of 

the results.
Non-specific  staining  as  a  result  of  unbound  probe  has  a  scattered,  granular  appearance 

and may be visualized at or distant from the expected hybridization site. Use intact cells for 

interpretation of staining results. Necrotic or degenerated cells may stain non-specifically (22).