Unit:
Κατεύθυνση Κλινική ΧημείαLibrary of the School of Science
Supervisors info:
J.Traeger-Synodinos Αναπλ. Καθηγήτρια
Original Title:
Σχεδιασμός και ανάπτυξη πρωτότυπου πρωτοκόλλου για γενετική διάγνωση μονογονιδιακού νοσήματος στην πρόληψη. Το παράδειγμα της α-Μεσογειακής αναιμίας.
Translated title:
Design and development of an original protocol for genetic diagnosis of monogenic disease in terms of prevention. The alpha-thalassemia example.
Summary:
Alpha-Thalassemia is a genetically inherited disease that occurs at very high
rates in both the Greek population and in the rest of the sub-tropic zone of
the planet. It is associated with mutations involving two genes (HBA1 and HBA2)
encoding two of the four polypeptide chains of hemoglobin. The phenotype of
humans carrying mutations in these genes varies. It ranges from conditions that
have almost no effect on the phenotype to situations incompatible with life, eg
Hydrops Fetalis. Therefore prevention in the case of carriers who wish to bear
children is indicated. In addition DNA analysis supports definitive diagnosis
of patients.
In this study we initially tried to optimize an existing protocol for direct
application in clinical practice as a screening method for point mutations in
the HBA1 and HBA2 genes based on High Resolution Melting Analysis (HRMA). Given
the fact that certain parameters in the experimental process were associated
with inconsistent results, it was decided to design and develop a new protocol.
The new protocol additionally had many advantages over the first, eg lower
cost, less steps and shorter time to result. In addition, we designed primers
to incorporate parts of the two genes, that were not included previously, as
well as a target specific probe for detecting a specific mutation that could
not clearly be separated from the wild type samples.
The results have been encouraging. Namely we detected without any doubt
mutations at any point of the targeted genes HBA1 and HBA2. Also, during the
conduct of experiments we detected two rare mutations in the Greek population
(Hb Fort Worth and codon 115 + CC), one of which was detected in the newly
designed parts of the genes.
High Resolution Melting Analysis (HRMA) was chosen as the detection method for
it is a quick, reliable and cost efficient technology, with several advantages
over existing methods, facts that make HRMA an excellent tool in any clinical
laboratory for point mutations.
Keywords:
HRMA, Prevention, Hb Fort Worth, Codon 115 (+CC), a-thalassemia
Number of index pages:
1-9
Number of references:
106
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