Abstract
Aims Manual grading is currently employed for HER-2 immuno-histochemistry (IHC). However, recent guidelines incorporated the use image analysisto reduce scoring variability.1 We utiliseda freely available image analysis programme to develop a semi-quantitative grading system for HER-2 IHC, validated it and compared it with manual grading and HER-2 gene amplification by CISH/SISH. Methods Initial testing was performed on 50 separate HER-2 immunohistochemically stained slides using ImageJ image analysis software v1.43 (NIH, Bethesda, MD, USA; http://rsb.in-fo.nih.gov/ij/). An image analysis grading system for HER-2 IHC (negative, equivocal and positive) was developed from the initial data. Another 60 cases were used for validation and comparison with manual grading and CISH/SISH (Pathology Department, Peter MacCallum Cancer Centre, Victoria). Results The concordance between the image analysis and the manual grading systems was 86.7% (k 0.79). Negative cases and positive cases on HER-2 IHC reached 98% and 100% concordance, respectively, with CISH/SISH for both the image analysis and the manual grading systems. However, the number of equivocal cases was reduced by 86% by image analysis when compared to manual grading. Discussion While both the image analysis and the manual grading have high sensitivity and specificity in predicting HER-2 gene amplification by CISH/SISH, image analysis can be utilised to reduce significantly the number of equivocal manually graded cases. Manual grading is currently employed for HER-2 immuno-histochemistry (IHC). However, recent guidelines incorporated the use image analysisto reduce scoring variability.1 We utiliseda freely available image analysis programme to develop a semi-quantitative grading system for HER-2 IHC, validated it and compared it with manual grading and HER-2 gene amplification by CISH/SISH. Initial testing was performed on 50 separate HER-2 immunohistochemically stained slides using ImageJ image analysis software v1.43 (NIH, Bethesda, MD, USA; http://rsb.in-fo.nih.gov/ij/). An image analysis grading system for HER-2 IHC (negative, equivocal and positive) was developed from the initial data. Another 60 cases were used for validation and comparison with manual grading and CISH/SISH (Pathology Department, Peter MacCallum Cancer Centre, Victoria). The concordance between the image analysis and the manual grading systems was 86.7% (k 0.79). Negative cases and positive cases on HER-2 IHC reached 98% and 100% concordance, respectively, with CISH/SISH for both the image analysis and the manual grading systems. However, the number of equivocal cases was reduced by 86% by image analysis when compared to manual grading. While both the image analysis and the manual grading have high sensitivity and specificity in predicting HER-2 gene amplification by CISH/SISH, image analysis can be utilised to reduce significantly the number of equivocal manually graded cases.
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