A comparison of Direct sequencing, Pyrosequencing, High resolution melting analysis, TheraScreen DxS, and the K-ras StripAssay for detecting KRAS mutations in non small cell lung carcinomas

• Published in: Journal of experimental & clinical cancer research 2012-09, Vol.31 (1), p.79-79, Article 79
• Main Authors: Jancik, Sylwia, Drabek, Jiri, Berkovcova, Jitka, Xu, Yong Zhong, Stankova, Marcela, Klein, Jiri, Kolek, Vitezslav, Skarda, Josef, Tichy, Tomas, Grygarkova, Ivona, Radzioch, Danuta, Hajduch, Marian
• Format: Article
• Language: English
• Subjects: Cancer; Carcinoma, Non-Small-Cell Lung - genetics; Care and treatment; Colorectal cancer; Colorectal Neoplasms - genetics; Colorectal Neoplasms - secondary; Cost-Benefit Analysis; Deoxyribonucleic acid; DNA; DNA sequencing; DNA, Neoplasm - analysis; DNA, Neoplasm - genetics; Epidermal growth factor; Genes; Genetic aspects; Genetic testing; Genetics; Genotyping; Humans; KRAS - Kiras2 kristen rat sarcoma viral oncogene homolog; Lung cancer; Lung cancer, Non-small cell; Medicine; Metastasis; Methods; Mutation; NSCLC - Non-small cell lung cancer; Nucleotide sequencing; Pathology; Patients; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins p21(ras); ras Proteins - genetics; Sensitivity and Specificity; Sequence Analysis, DNA - economics; Sequence Analysis, DNA - methods; Single nucleotide polymorphisms; SNP - single nucleotide polymorphism; Studies; Surgery
• ISSN: 1756-9966; 0392-9078; 1756-9966
• DOI: 10.1186/1756-9966-31-79

It is mandatory to confirm the absence of mutations in the KRAS gene before treating metastatic colorectal cancers with epidermal growth factor receptor inhibitors, and similar regulations are being considered for non-small cell lung carcinomas (NSCLC) and other tumor types. Routine diagnosis of KRAS mutations in NSCLC is challenging because of compromised quantity and quality of biological material. Although there are several methods available for detecting mutations in KRAS, there is little comparative data regarding their analytical performance, economic merits, and workflow parameters. We compared the specificity, sensitivity, cost, and working time of five methods using 131 frozen NSCLC tissue samples. We extracted genomic DNA from the samples and compared the performance of Sanger cycle sequencing, Pyrosequencing, High-resolution melting analysis (HRM), and the Conformité Européenne (CE)-marked TheraScreen DxS and K-ras StripAssay kits. Our results demonstrate that TheraScreen DxS and the StripAssay, in that order, were most effective at diagnosing mutations in KRAS. However, there were still unsatisfactory disagreements between them for 6.1% of all samples tested. Despite this, our findings are likely to assist molecular biologists in making rational decisions when selecting a reliable, efficient, and cost-effective method for detecting KRAS mutations in heterogeneous clinical tumor samples.