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Identifier

000419777

Title

Detection of BRAFV600E mutation via a diagnostic system based on acoustic biosensors

Alternative Title

Ανίχνευση της μετάλλαξης BRAFV600E μέσου ενός διαγνωστικού συστήματος βασισμένου σε ακουστικούς βιοαισθητήρες.

Author

Θεοδόση, Γεσθημανή-Ιωάννα Δ.

Thesis advisor

Γκιζελή, Ηλέκτρα
Παπαδάκης, Γιώργος

Abstract

Diagnostic tests for detection of mutations in cancer-related genes are of great importance for cancer control and management. In this work, we introduce a new methodology based on isothermal amplification and acoustic biosensors for the detection of BRAF^V600E mutation, which is very common in melanoma patients and it is the target of specific therapeutic approaches. More specifically, we tried to develop an assay that could have the potential to be applied in both solid and liquid biopsy. Thus, we employed both genomic DNA and DNA fragments as templates for loop-mediated isothermal amplification (LAMP) and quartz crystal microbalance (QCM) technology for the biosensor-based detection. The acoustic measurements were expressed as ratio of dissipation versus frequency change (ΔD/ΔF). However, the percentages of mutant ratio relative to wild-type, show clearer differences. Interestingly, a positive correlation between them and mutant allele frequency (MAF) was observed in case of genomic DNA. Peptide nucleic acid (PNA) was utilized to reduce the wild-type background. Finally, through the proposed method we managed to detect mutant allele with a frequency of 5% in genomic DNA and 0.1% in fragments, comparing every single mutant to its corresponding wild-type prepared with the same set of materials. The described assay holds promise to serve as a new simple, fast and cost-effective diagnostic solution that could contribute to an effective and safe treatment guidance for patients.

Language

English

Subject

Diagnostics

Melanoma

Διαγνωστικά

Μελάνωμα

Issue date

2018-11-23

Collection

School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses

Type of Work--Post-graduate theses