title: EGFR activating mutations detected by different PCR techniques in Caucasian NSCLC patients with CNS metastases: short report.
abstract: EGFR mutation testing has become an essential determination to decide treatment options for NSCLC. The mutation analysis is often conducted in samples with low percentage of tumour cells from primary tumour biopsies. There is very little evidence that samples from metastatic tissues are suitable for EGFR testing. We had evaluated the frequency of EGFR mutations with three highly sensitive PCR techniques in formalin-fixed, paraffin-embedded samples of 143 NSCLC patients with central nervous system (CNS) metastases. 32 corresponding primary tumours were also examined. We used PCR followed by DNA fragments length analysis (FLA), ASP-PCR and PNA-LNA PCR clamp techniques. We found 9 (6.29 %) EGFR gene mutations in CNS samples: 3 (2.1 %) in exon 19 and 6 (4.2 %) in exon 21. The full concordance between CNS metastases and primary tumour samples was observed. PCR followed by DNA-FLA and PNA-LNA PCR clamp were sensitive enough to detect exon 19 deletions. Two mutations in exon 21 were detected by ASP-PCR only, one L858R substitution was detected only by PNA-LNA PCR clamp. With respect to sensitivity, PCR followed by DNA-FLA achieved a level of detection of at least 10 % of mutated DNA for exon 19 deletion, as for ASP-PCR it was at least 5 % of mutated DNA for L858R substitution. Higher sensitivity of 1 % of mutated DNA was achieved by PNA-LNA PCR clamp technique for both mutations. The use of different methodological techniques authenticates the negative result of molecular tests.
authors.affiliation: Pneumonology, Oncology and Allergology Department, Medical University of Lublin, Jaczewskiego 8, 20-954, Lublin, Poland, kamilawojas@wp.pl.
authors.forename: Wojas-Krawczyk
authors.initials: WK
authors.lastname: Kamila
authors.forename: Skroński
authors.initials: S
authors.lastname: Michał
authors.forename: Krawczyk
authors.initials: K
authors.lastname: Paweł
authors.forename: Jaguś
authors.initials: J
authors.lastname: Paulina
authors.forename: Kucharczyk
authors.initials: K
authors.lastname: Tomasz
authors.forename: Jarosz
authors.initials: J
authors.lastname: Bożena
authors.forename: Mlak
authors.initials: M
authors.lastname: Radosław
authors.forename: Szumiło
authors.initials: S
authors.lastname: Justyna
authors.forename: Sawicki
authors.initials: S
authors.lastname: Marek
authors.forename: Tomasz
authors.initials: T
authors.lastname: Trojanowski
authors.forename: Milanowski
authors.initials: M
authors.lastname: Janusz
authors.forename: Chorostowska-Wynimko
authors.initials: CW
authors.lastname: Joanna
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology; D000368:Aged; D001932:Brain Neoplasms / Q000235:genetics* / Q000556:secondary; D018287:Carcinoma, Large Cell / Q000235:genetics / Q000473:pathology; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000473:pathology; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000473:pathology; D004273:DNA, Neoplasm / Q000235:genetics*; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics*; D009367:Neoplasm Staging; D016133:Polymerase Chain Reaction / Q000379:methods*; D011379:Prognosis; D012307:Risk Factors
chemical_list: D004273:DNA, Neoplasm; C512478:EGFR protein, human; D066246:ErbB Receptors
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