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Application of image analysis in external and internal quality assurance for diagnostic clinical immunohistochemistry2012 October 1900 (has links)
Clinical immunohistochemistry (IHC) techniques are not yet fully standardized. In this project, a standardization method was developed and tested for proficiency testing (PT) in external quality assurance (EQA) and quality control (QC) in clinical IHC laboratories. The breast cancer markers estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor 2 (HER2) were used as a model system. Digital image analysis (IA) was used in conjunction with new calibrated and standardized cell line microarrays (CLMA). CLMAs built from nine formalin-fixed paraffin-embedded (FFPE) breast cancer cell lines were used for both QC controls and PT samples, instead of traditionally used FFPE tissues, in the standardization of breast cancer IHC. IA was used for measurement of IHC results, and compared to evaluation by the traditional expert-assessment method.
Laboratory Score: Reference Score Ratio (LSRSR) was derived from Histo-Scores (HScores) determined by IA. HScores and LSRSRs were examined statistically and evaluated as histograms and boxplots to summarize and rank participant laboratory EQA results, in comparison to a reference sample or reference laboratories in two consecutive Canada-wide EQA runs.
LSRSR-derived reference ranges were highly sensitive in evaluating laboratory EQA performance in PT as well as for monitoring of controls for QC. Laboratory on-slide tissue and cell-line IHC QA controls were assessed using IA and Levey Jennings QC charts. These charts were determined to be an excellent way to observe trending in laboratory IHC staining over time, particularly when cell line controls were used. This approach also reduced the time and labor costs for PT evaluation.
Overall, cell line calibration controls were functionally equivalent or better than tissue-based controls in QC and PT mainly because of cell line biological homogeneity and sample availability. This study identified an optimal design for preparation of IHC cell line controls and PT samples for breast cancer markers. Optimal, intermediate staining cell line IHC controls were identified for all three breast cancer markers.
Using IA with LSRSR and cell line samples is recommended for standardization of IHC methodology. This approach advances QA for diagnostic IHC and when implemented will improve patient care
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