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M. Levi
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P3.01 - Advanced NSCLC (ID 621)
- Event: WCLC 2017
- Type: Poster Session with Presenters Present
- Track: Advanced NSCLC
- Presentations: 1
- Moderators:
- Coordinates: 10/18/2017, 09:30 - 16:00, Exhibit Hall (Hall B + C)
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P3.01-053 - Detection of Common EGFR Mutation in Cytological Smears Using Reversed Dot Blot (RDB) Hybridization Method (ID 9765)
09:30 - 09:30 | Author(s): M. Levi
- Abstract
Background:
Epidermal growth factor receptor (EGFR) mutation can predict the response to EGFR tyrosine kinase inhibitor (TKI) in non small cell lung cancer (NSCLC). In this study, we determined the limit of detection of different methods, such as sequencing, restriction frament length polymorphism (RFLP), high resolution melting (HRM) and reversed dot blot (RDB) hybridization to detect for EGFR mutation detection.
Method:
Mutation detections of exons19 and 21 in EGFR gene were performed using sequencing, restriction fragment length polymorphism (RFLP) and high resolution melting (HRM) analysis method. Moreover, we also developed Reversed Dot Blot(RDB) method hybridization as an alternative procedure. Genomic DNA of H1975, HCT116 cell line as well as specially designed oligonucleotides were used to determine the limit of detection of these procedures. We also determine the sensitivity and specificity of RDB method compared to existing methods.
Result:
Limit of detection of each method was determined by titration of mutant allele in wildtype background. HRM analysis and RFLP were able to detect mutation in samples containing 6.25% to 12.5% mutated DNA.Limit of detection of RDB method was around 12.5%. Analysis of 40 patients had shown that the specificity for to detect mutations in exon 19 and 21 was 100% and 89.5% and the sensitivity for both exon was 62.5%.
Conclusion:
RDB method maybe used as an alternative method to standard procedure such as sequencing. This method is specific, but need further improvement to increase its sensitivity.
