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EGFR mutation detection by microfluidic technology: a validation study
  1. Umberto Malapelle1,
  2. Stefania Russo1,
  3. Francesco Pepe1,
  4. Roberta Sgariglia1,
  5. Caterina De Luca1,
  6. Claudio Bellevicine1,
  7. Pierlorenzo Pallante2,
  8. Giancarlo Troncone1
  1. 1Dipartimento di Sanità Pubblica, Università di Napoli Federico II, Napoli, Italia
  2. 2Consiglio Nazionale delle Ricerche, Istituto di Endocrinologia ed Oncologia Sperimentale, Napoli, Italia
  1. Correspondence to Professor Giancarlo Troncone, Dipartimento di Sanità Pubblica, Università di Napoli Federico II, via Sergio Pansini 5, Napoli I-80131, Italy; giancarlo.troncone{at}unina.it

Abstract

Advanced non-small cell lung cancer samples are tested for epidermal growth factor receptor (EGFR) gene mutations. Their detection by direct sequencing is time-consuming. Conversely, the length analysis of fluorescently labelled PCR products is easier. To avoid labelled primers and the automated capillary electrophoresis apparatus, we validated a fast and sensitive chip-based microfluidic technology. The limit of detection of fragment length assay on microfluidic device was 5%, more sensitive than direct sequencing (12.5%). The novel methodology showed high accuracy in the analysis of samples whose mutational status was known. The accuracy in quantifying mutated alleles (mA) was evaluated by PCR products subcloning; the mA% provided by direct sequencing of subcloned PCR products showed a close correlation with the mA% provided by the microfluidic technology for both exon 19 (R2=0.9) and 21 (R2=0.9). Microfluidic-based on-chip electrophoresis makes EGFR testing more rapid, sensitive and cost-effective.

  • EGFR
  • LUNG
  • MOLECULAR PATHOLOGY

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