Comparison of Four Different Scoring Methods for Ki-67 Percentage in Breast Carcinoma: A Cross-sectional Study
Authors: Angel Evelyn , Muralee Praseeda C , Mathew Sherin Susheel , Bhasy Sreeja K , Roselin . , Kumar V Dinesh , Nasar Anisha , Panicker Priya 
Published: May 2024

Introduction: In breast cancer, the prognostic role of Ki-67 has been comprehensively studied, and its usefulness has been proven. Ki-67 expression has a prognostic and predictive value in both adjuvant therapy response and neoadjuvant settings. However, the inter and the intraobserver variability in manual counting limits the accuracy of scoring Ki-67 and consequently its application in treatment. Aim: To examine four different methods of Ki-67 estimation to find the most reliable, reproducible and time-efficient scoring methods for Ki-67. Materials and Methods: The present cross-sectional study was conducted in the Department of Pathology, Sree Mookambika Institute of Medical Sciences, Nagercoil, Tamil Nadu, India, from May 2023 to August 2023. Ki-67 immunostained slides of 30 trucut biopsies of invasive breast carcinoma were retrieved and analysed by two observers in a blinded manner. The four different methods of analysis of Ki-67 expression carried out were the global method, the hotspot method, the Eye-10 method and the stepwise counting strategy. The parameters included in the present study were the mean age of the study population, the pre/postmenopausal status, the histopathological type of invasive breast carcinoma, Estrogen receptor (ER), Progesterone Receptor (PR), and Human Epidermal growth factor Receptor 2 (HER2)/neu status, as well as, the interobserver agreement and the mean time taken by the four methods to analyse the Ki-67 expression. Data entry was performed using Microsoft Office Excel 2013, and statistical analysis was conducted using Statistical Package for the Social Sciences (SPSS) software version 20.0. Descriptive statistics for qualitative variables and mean and Standard Deviation (SD) for quantitative variables were used for data analysis. Chi-square tests were used for bivariate analysis with a determined statistical significance of 5% (p-value<0.05). Results: The mean age of the study population was 52.9±9.1 years. The highest interobserver concordance was observed among the observers using the weighted global scoring method, with an Interclass Correlation Coefficient (ICC) of 0.967. This method was time consuming, with the first observer taking a mean±SD time of 5.5±0.8 minutes and the second observer taking 4.6±0.9 minutes. The least time was consumed for performing the Eye-10 scoring, with mean±SD time was 1±0.87 minutes. The stepwise counting and hotspot method demonstrated excellent inter-rater agreement, with a kappa of 0.8304 (p-value<0.001) between both observers. Conclusion: Although the Eye-10 method and stepwise counting took the least time, they are limited by a gray/intermediate zone for scoring compared to the global scoring method and the hotspot method. As a result, the global and/or hotspot method after proper training is a robust and reliable method of assessing Ki-67, with the hotspot method being the most reliable, as the global method is limited by the use of an online tool.