Optimization of routine KRAS mutation PCR‐based testing procedure for rational individualized first‐line‐targeted therapy selection in metastatic colorectal cancer

Cancer Medicine

Anne-Sophie Chretien²  Alexandre Harlé³  Magali Meyer-Lefebvre²  Marie Rouyer²  Marie Husson²  Carole Ramacci²  Valentin Harter¹  Pascal Genin²  Agnès Leroux² 

[1] Service de Biostatistiques, Centre Alexis Vautrin, 6 Avenue de Bourgogne, Vandœuvre-lès-Nancy, France;Service de Biopathologie, Centre Alexis Vautrin, Vandœuvre-lès-Nancy, France;CNRS, UMR 7039 CRAN, Nancy, France

关键词: Colorectal cancer;    HRM;    KRAS;    PCR‐RFLP;    TaqMan PCR;   

DOI  :  10.1002/cam4.47

来源: Wiley

【 摘 要 】

Abstract

KRAS mutation detection represents a crucial issue in metastatic colorectal cancer (mCRC). The optimization of KRAS mutation detection delay enabling rational prescription of first-line treatment in mCRC including anti-EGFR-targeted therapy requires robust and rapid molecular biology techniques. Routine analysis of mutations in codons 12 and 13 on 674 paraffin-embedded tissue specimens of mCRC has been performed for KRAS mutations detection using three molecular biology techniques, that is, high-resolution melting (HRM), polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), and allelic discrimination PCR (TaqMan PCR). Discordant cases were assessed with COBAS 4800 KRAS CE-IVD assay. Among the 674 tumor specimens, 1.5% (10/674) had excessive DNA degradation and could not be analyzed. KRAS mutations were detected in 38.0% (256/674) of the analysable specimens (82.4% in codon 12 and 17.6% in codon 13). Among 613 specimens in whom all three techniques were used, 12 (2.0%) cases of discordance between the three techniques were observed. 83.3% (10/12) of the discordances were due to PCR-RFLP as confirmed by COBAS 4800 retrospective analysis. The three techniques were statistically comparable (κ > 0.9; P < 0.001). From these results, optimization of the routine procedure consisted of proceeding to systematic KRAS detection using HRM and TaqMan and PCR-RFLP in case of discordance and allowed significant decrease in delays. The results showed an excellent correlation between the three techniques. Using HRM and TaqMan warrants high-quality and rapid-routine KRAS mutation detection in paraffin-embedded tumor specimens. The new procedure allowed a significant decrease in delays for reporting results, enabling rational prescription of first-line-targeted therapy in mCRC.

【 授权许可】

CC BY   
© 2012 The Authors. Cancer Medicine published by Blackwell Publishing Ltd.

Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.