Ki-67 Proliferation Index in Breast Cancer as a Function of Assessment Method: A NordiQC Experience.

Immunohistochemical staining for Ki-67 is used to calculate a Ki-67 proliferation index (PI) that carries prognostic and predictive information in various cancers including breast carcinomas. Studies have documented challenges for observers to reproducibly estimate the Ki-67 PI. At present, no international consensus exists concerning scoring method (eg, hotspots vs. overall average, digital vs. manual counting) or even the definition of a Ki-67-positive cell. To clarify the approach to Ki-67 scoring and evaluation of the interobserver agreement among participants in the Nordic Immunohistochemical Quality Control (NordiQC) Breast Cancer Module, a study was set up on the basis of an online web module containing 15 digitized tissue microarray cores of breast carcinomas stained for Ki-67 in the NordiQC reference laboratory. All participants were invited to attend the study. In addition to Ki-67 scoring, they were asked to disclose their preferred method for Ki-67 estimation and their job title. For comparison, slides were analyzed using a Digital Image Analysis algorithm based on Virtual Double Staining. In total, 199 participants enrolled for the study. Overall, there was a good correlation in Ki-67 PIs among the participants, although results for some cores varied significantly. However, when applying a cutoff of 20%, a relatively low kappa value of 0.52 was observed. Participants scoring in hotspots reported higher Ki-67 PIs than participants estimating an overall average, and, not surprisingly, participants who considered weak Ki-67 nuclear staining positive obtained higher Ki-67 PIs than those who did not. Ki-67 PI was not correlated to job title. The Virtual Double Staining algorithm obtained Ki-67 values close to the mean value of the human observers. Our study underlines the need for international standardization and guidelines in estimation of Ki-67 PI. Digital Image Analysis may be a useful tool in this process.