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| Identifier |
000399221 |
| Title |
Detection of KRAS gene mutations by PNA-based qPCR in plasma cell-free DNA of colorectal cancer patients |
| Alternative Title |
Ανίχνευση μεταλλάξεων του γονιδίου KRAS με τη χρήση PNA-based qPCR στο κυκλοφορούν DNA του πλάσματος ασθενών με καρκίνο του παχέος εντέρου |
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Author
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Γκουντάκος, Αναστάσιος Ι.
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Thesis advisor
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Μαυρουδής, Δ.
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Reviewer
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Βενυχάκη, Μ.
Σουρβίνος, Γ.
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| Abstract |
Introduction: Colorectal cancer (CRC) is one of the leading causes of cancer-related death in the Western world while the mortality and survival rates are highly influenced by the stage of disease at diagnosis. Several reports have demonstrated the existence of elevated cell-free DNA levels in the circulation of CRC patients, suggesting the possibility to develop and validate a reliable non-invasive genetic biomarker. The purpose of this study was to investigate the presence of KRAS mutations in plasma cell-free DNA from patients with CRC.
Materials and Methods: Cell-free DNA was extracted by QIAamp CNA kit and serum and plasma cell-free DNA levels were quantified by quantitative real-time PCR. The identification of three of the most common KRAS mutations on codon 12 and 13 was performed by Peptide Nucleic Acid (PNA)-based qPCR assay.
Results: Our observations confirmed the significance of the 2-step centrifugation during the plasma isolation. The data of this study also demonstrated the presence of significantly higher serum and plasma cfDNA levels in CRC patients compared to healthy individuals (control group). Moreover, the mutation analysis in the plasma cfDNA revealed detectable KRAS mutations in 5/9 (55%) pre-chemotherapy samples and in 8/10 (80%) post-chemotherapy samples.
Conclusions: Our results suggest that circulating cell-free DNA detectable with a well-known and validated qPCR assay could represent an innovative, minimally invasive approach to detect and monitor RAS mutation status in CRC patients.
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| Language |
English |
| Subject |
Colorecta cancer |
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Serum |
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Βιοδείκτης |
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Ορός |
| Issue date |
2015-07-17 |
| Collection
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School/Department--School of Medicine--Department of Medicine--Post-graduate theses
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Type of Work--Post-graduate theses
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| Views |
170 |
Detection of KRAS gene mutations by PNA-based qPCR in plasma cell-free DNA of colorectal cancer patients
