EGFR mutation detection by microfluidic technology: a validation study.

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 (R(2)=0.9) and 21 (R(2)=0.9). Microfluidic-based on-chip electrophoresis makes EGFR testing more rapid, sensitive and cost-effective.

journal_name

J Clin Pathol

authors

Malapelle U,Russo S,Pepe F,Sgariglia R,De Luca C,Bellevicine C,Pallante P,Troncone G

doi

10.1136/jclinpath-2013-201730

subject

Has Abstract

pub_date

2013-11-01 00:00:00

pages

982-4

issue

11

eissn

0021-9746

issn

1472-4146

pii

jclinpath-2013-201730

journal_volume

66

pub_type

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