A Highly Verified Assay for KRAS Mutation Detection in Tissue and Plasma of Lung, Colorectal, and Pancreatic Cancer

• Published in: Archives of pathology & laboratory medicine (1976) Vol. 143; no. 2; pp. 183 - 189
• Main Authors: Li, Jing, Gan, Stephanie, Blair, Alan, Min, Kyungji, Rehage, Taraneh, Hoeppner, Corey, Halait, Harkanwal, Brophy, Victoria H.
• Format: Journal Article
• Language: English
• Published: United States College of American Pathologists 01.02.2019
• Subjects: Biochemical assays; Biomarkers; Body fluids; Cancer patients; Cancer screening; Clinical medicine; Colon cancer; Colorectal cancer; Colorectal Neoplasms - genetics; Deoxyribonucleic acid; Diagnosis; DNA; DNA Mutational Analysis - methods; EDTA; Formaldehyde; Gene mutation; Genes; Genetic aspects; Health aspects; Humans; Identification and classification; Initiatives; K-Ras protein; Laboratories; Lung cancer; Lung Neoplasms - genetics; Medical screening; Metastases; Metastasis; Mutation; Non-small cell lung cancer; Oncology; Operators; Pancreatic cancer; Pancreatic Neoplasms - genetics; Plasma; Proto-Oncogene Proteins p21(ras) - genetics; Real-Time Polymerase Chain Reaction - methods; Regulatory genes; Sensitivity and Specificity; Small cell lung cancer; Testing; Therapeutic applications; Tumors; Working groups
• ISSN: 0003-9985; 1543-2165; 1543-2165
• DOI: 10.5858/arpa.2017-0471-OA

KRAS Mutation Test v2 is used for the qualitative detection and identification of 28 mutations in exons 2, 3, and 4 of the human KRAS gene. To verify the performance of KRAS Mutation Test v2 and to evaluate its accuracy by correlation with a next-generation sequencing method on Illumina MiSeq. In this study, we used formalin-fixed, paraffin-embedded tissue and plasma specimens from non-small cell lung cancer, colorectal cancer, and pancreatic cancer patients. Results of specificity, precision, analytical sensitivity, and accuracy as compared with a MiSeq method are reported. The KRAS Mutation Test v2 demonstrated exquisite sensitivity and specificity and broad coverage of KRAS mutations. Precision was 100% (108 of 108) across all samples, operators, and instruments for formalin-fixed, paraffin-embedded tissue and 99.8% (615 of 616) for plasma. Analytical sensitivity was high with detection of 1% mutant sequence in formalin-fixed, paraffin-embedded tissue samples and as low as 25 mutant sequence copies/mL for plasma samples. The test also showed high overall concordance for formalin-fixed, paraffin-embedded tumor tissue as well as for plasma specimens when compared with MiSeq sequencing results. The KRAS Mutation Test v2 is a highly robust, reproducible, and sensitive test for the qualitative detection of 28 mutations in exons 2, 3, and 4 of the KRAS gene in both solid (tissue) and liquid (plasma) biopsies from colorectal cancer, non-small cell lung cancer, and pancreatic cancer, and is a convenient option for KRAS mutation testing.