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- W1959178307 abstract "Reliable detection of EGFR mutations in lung cancer metastases requires highly sensitive and robust molecular method. Our aim was to assess effectiveness of two highly sensitive PCR methods in detection of EGFR activating mutations in FFPE samples of brain metastases from 142 NSCLC patients. Isolated DNA was analyzed for EGFR exon 19 deletions and exon 21 L858R mutations by real-time PCR PNA-LNA PCR clamp and allele-specific PCR (ASP-PCR). If discrepant, results were re-evaluated by TaqMan genotyping. Direct sequencing analyses are ongoing. All samples were successfully analyzed. In 37 samples (26%) DNA was of low quality and PCR pre-amplification was performed prior to PNA-LNA PCR clamp analysis. In total 11 out of 142 samples (8%) proved positive for EGFR activating mutations. 6 samples (55% of detected) were positive for exon 19 deletions (n=3) and L858R substitution (n=3) as assessed by both PNA-LNA PCR clamp and ASP-PCR. ASP-PCR, but not PNA-LNA PCR clamp, detected 3 further L858R mutations, whereas 1 L858R substitution and 1 rare A859T mutation were detected by PNA-LNA PCR clamp only. None of discrepant L858R substitutions were confirmed by TaqMan genotyping. Different approach to molecular diagnostics represented by two highly sensitive methods employed in our study might be responsible for observed discrepancies. ASP-PCR utilizes allele-specific primers while in PNA-LNA PCR clamp method PNA probe inhibits wild type allele amplification. In FFPE samples with low tumor cells content, where direct sequencing is not applicable, it might be advisable to re-evaluate diagnostic results by another equally sensitive, but methodologically different diagnostic technique." @default.
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- W1959178307 date "2012-09-01" @default.
- W1959178307 modified "2023-09-27" @default.
- W1959178307 title "Reliability of EGFR mutations detection in NSCLC brain metastases by two different allele-specific PCR methods" @default.
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