RT Journal Article
SR Electronic
T1 EGFR mutation detection on routine cytological smears of non-small cell lung cancer by digital PCR: a validation study
JF Journal of Clinical Pathology
JO J Clin Pathol
FD BMJ Publishing Group Ltd and Association of Clinical Pathologists
SP 454
OP 457
DO 10.1136/jclinpath-2015-203429
VO 69
IS 5
A1 Umberto Malapelle
A1 Caterina de Luca
A1 Elena Vigliar
A1 Francesca Ambrosio
A1 Danilo Rocco
A1 Pasquale Pisapia
A1 Claudio Bellevicine
A1 Giancarlo Troncone
YR 2016
UL http://jcp.bmj.com/content/69/5/454.abstract
AB Highly sensitive genotyping techniques are useful to detect epidermal growth factor receptor (EGFR) mutations on lung cancer cytological samples, when these specimens feature only few neoplastic cells. This study aimed to validate digital PCR (dPCR) methodology on cytological material. In plasmid model system, dPCR allowed for the detection of a minimal percentage (1%) of EGFR mutant alleles. Cytological samples (n=30), with neoplastic cell percentage ranging from 10% to 80% and yielding a quantity of extracted DNA ranging from 1.75 to 60 ng/µL were selected. Results previously generated by fragment length and TaqMan assays (n=8 exon 19 deletions, n=2 L858R mutations and n=20 wild-type DNA) were compared with those obtained by dPCR. Data were highly concordant (96.6%). However, dPCR detected an additional L858R mutation that had been missed by TaqMan assay on a paucicellular smear. This mutation was confirmed by cloning PCR products and sequencing. Thus, dPCR can reliably be used to increase EGFR mutation detection rate on scarcely cellular lung cancer smears.