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Identifier

000412656

Title

Detection of Circulating Nucleic Acids associated with cancer using QCM-D acoustic device combined with RCA isothermal DNA amplification method

Alternative Title

Ανίχνευση κυκλοφορούντων νουκλεϊκών οξέων που σχετίζονται με τον καρκίνο με τη χρήση ακουστικών βιοαισθητήρων σε συνδυασμό με την ισόθερμη μέθοδο πολλαπλασιασμού του DNA RCA

Author of screenplay

Ναούμη, Νιικολέττα Χ.

Thesis advisor

Γκιζέλη, Ηλέκτρα

Abstract

Cancer is the leading cause of death worldwide. The gold standards of cancer diagnosis are radiology, magnetic resonance imaging (MRI) and solid biopsies. However all three methods are expensive, time-consuming and hurt the physical health of patients during the detection of cancer. Circulating tumor DNA and circulating miRNA, both cancer biomarkers, have been widely used as targets for the development of non-invasive diagnostic methods for prognosis and diagnosis of cancer. A big variety of molecular, amplification-based techniques as well as methods that utilize the advances in nanomaterilas, nanotechnology and the biosensors field have been developed, too. However, most of these methods are expensive, complex, require special instrumentation, and a lot of time, so they cannot be applicable in routine clinical practice. Herein, it was aimed the development of two non-inavnsive, easy and fast methods for the detection of ctDNA point mutations and circulating miRNAs. Both methods utilize the QCM-D acoustic biosensor, combined with the isothermal DNA amplification method, RCA. In the case of ctDNA, a minimal amount of 10 copies was detected with high specificity via Ligation followed by RCA, but only 1 pmol was managed to be detected on QCM biosensor. Concerning the second case, 1 fmol of miRNA-21 was achieved to be detected off-chip in a single-step reaction (combining the ligation and amplification step in one single step reaction) at RT, without the need of any kind of instrumentation. However, only 0.1 pmol of miRNA-21 was detected on QCM-D device. Despite both methods need a lot of further optimization, results were promising for the development of a novel detection method.

Language

English

Subject

Acoustic Biosensors

Ακουστικοί βιοαισθητήρες

Κυκλοφορούντα νουκλεϊκά οξέα

Issue date

2017-11-22

Collection

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

Type of Work--Post-graduate theses

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