Performance Evaluation Comparison of Three Commercially Available PCR-Based KRAS Mutation Testing Platforms
Julia A Adams, Kristin M Post, Sarah A Bilbo, Xiaoyan Wang, Joyashree D Sen, Anita J Cornwell, Amanda J Malek, Liang Cheng. Indiana University School of Medicine, Indianapolis, IN
Background: The identification of KRAS mutations in patients with certain types of cancer, including colonic adenocarcinoma and non-small cell lung carcinoma has become increasingly important as these patients are contraindicated from receiving EGFR-targeted therapies. Several PCR-based tests are commercially available for KRAS mutation testing including Applied Biosystems KRAS Mutation Analysis on the ABI 3130xl, Qiagen therascreen KRAS RGQ PCR on the Rotor-Gene Q MDx and Qiagen KRAS Pyro on the PyroMark Q24, however these tests have not been compared side by side. The purpose of this study was to evaluate the performance characteristics and workflow for three PCR-based methods of detecting KRAS mutation status.
Design: We performed three of the commercially available PCR-based techniques for detecting the KRAS mutation, Applied Biosystems KRAS Mutation Analysis on the ABI 3130xl, Qiagen therascreen KRAS RGQ PCR on the Rotor-Gene Q MDx and Qiagen KRAS Pyro on the PyroMark Q24, on 188 patient samples and Acrometrix standards. The patient samples were of varying tissue types including colorectal, pancreatic, lung, omentum, and diaphragm. The results of each test for every specimen were documented and the three techniques were evaluated for sensitivity, specificity, positive predictive value, negative predictive value, accuracy, workflow, and cost.
Results: All of the 188 samples run were successful, with 29% being positive for the KRAS mutation. Of the positive tests, Applied Biosystems detected 84% of the positive cases, whereas Therascreen RGQ and Pyro detected 100% of the positive cases. In cases of discrepancy between Applied Biosystems and Therascreen RGQ, Pyro agreed with Therascreen RGQ 95% of the time. Therascreen RGQ and Pyro, were comparable in terms of sensitivity, specificity, positive predictive value, negative predictive value, and accuracy, with all values being 100%. All three techniques accurately identified the appropriate mutation in the known control specimens.
Conclusions: All three tests are relatively comparable for detecting the KRAS mutation, with Applied Biosystems having a slightly lower sensitivity, negative predictive value, and accuracy than Therascreen RGQ and Pyro.
Wednesday, March 6, 2013 9:30 AM
Poster Session V # 257, Wednesday Morning