Detection of EGFR mutations by TaqMan mutation detection assays powered by competitive allele-specific TaqMan PCR technology.

Abstract

Epidermal growth factor receptor (EGFR) mutations in non-small-cell lung cancer (NSCLC) are predictive of response to treatment with tyrosine kinase inhibitors. Competitive Allele-Specific TaqMan PCR (castPCR) is a highly sensitive and specific technology. EGFR mutations were assessed by TaqMan Mutation Detection Assays (TMDA) based on castPCR technology in 64 tumor samples: a training set of 30 NSCLC and 6 colorectal carcinoma (CRC) samples and a validation set of 28 NSCLC cases. The sensitivity and specificity of this method were compared with routine diagnostic techniques including direct sequencing and the EGFR Therascreen RGQ kit. Analysis of the training set allowed the identification of the threshold value for data analysis (0.2); the maximum cycle threshold (Ct = 37); and the cut-off ΔCt value (7) for the EGFR TMDA. By using these parameters, castPCR technology identified both training and validation set EGFR mutations with similar frequency as compared with the Therascreen kit. Sequencing detected rare mutations that are not identified by either castPCR or Therascreen, but in samples with low tumor cell content it failed to detect common mutations that were revealed by real-time PCR based methods. In conclusion, our data suggest that castPCR is highly sensitive and specific to detect EGFR mutations in NSCLC clinical samples.

• Authors: Cristin Roma, Claudia Esposito, Anna Maria Rachiglio, Raffaella Pasquale, Alessia Iannaccone, Nicoletta Chicchinelli, Renato Franco, Rita Mancini, Salvatore Pisconti, Antonella De Luca, Gerardo Botti, Alessandro Morabito, Nicola Normanno
• Journal: BioMed research international (Biomed Res Int) Vol. 2013 Pg. 385087 ( 2013) ISSN: 2314-6141 [Electronic] United States
• PMID: 24364033 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• EGFR protein, human
• ErbB Receptors

Topics

• Alleles
• Carcinoma, Non-Small-Cell Lung (genetics, pathology)
• Colorectal Neoplasms (genetics, pathology)
• ErbB Receptors (genetics, isolation & purification)
• Humans
• Mutation
• Polymerase Chain Reaction