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

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

Research output: Contribution to journalArticle

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 (C t = 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.

Original languageEnglish
Article number385087
JournalBioMed Research International
Volume2013
DOIs
Publication statusPublished - 2013

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Epidermal Growth Factor Receptor
Assays
Alleles
Technology
Polymerase Chain Reaction
Mutation
Non-Small Cell Lung Carcinoma
Tumors
Cells
Protein-Tyrosine Kinases
Real-Time Polymerase Chain Reaction
Colorectal Neoplasms
Neoplasms
Sensitivity and Specificity

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Detection of EGFR mutations by TaqMan mutation detection assays powered by competitive allele-specific taqman PCR technology. / Roma, Cristin; Esposito, Claudia; Rachiglio, Anna Maria; Pasquale, Raffaella; Iannaccone, Alessia; Chicchinelli, Nicoletta; Franco, Renato; Mancini, Rita; Pisconti, Salvatore; De Luca, Antonella; Botti, Gerardo; Morabito, Alessandro; Normanno, Nicola.

In: BioMed Research International, Vol. 2013, 385087, 2013.

Research output: Contribution to journalArticle

Roma, Cristin ; Esposito, Claudia ; Rachiglio, Anna Maria ; Pasquale, Raffaella ; Iannaccone, Alessia ; Chicchinelli, Nicoletta ; Franco, Renato ; Mancini, Rita ; Pisconti, Salvatore ; De Luca, Antonella ; Botti, Gerardo ; Morabito, Alessandro ; Normanno, Nicola. / Detection of EGFR mutations by TaqMan mutation detection assays powered by competitive allele-specific taqman PCR technology. In: BioMed Research International. 2013 ; Vol. 2013.
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