|
Identifier |
000452159 |
Title |
Detection of Sickle Cell Disease point-mutation for application at the point-of -care |
Alternative Title |
Ανίχνευση της σημειακής μετάλλαξης της δρεπανοκυτταρικής αναιμίας για εφαρμογή στο χώρο παροχής φροντίδας |
Author
|
Δημητριάδη, Άννα Ε.
|
Thesis advisor
|
Γκιζελή, Ηλέκτρα
|
Reviewer
|
Στρουμπούλης, Ιωάννης
Καρδάσης, Δημήτρης
|
Abstract |
Haemoglobinopathies refer to a diverse group of inherited disorders characterized
by a reduced synthesis of one or more globin chains (thalassemia) or the synthesis of
a structurally abnormal hemoglobin (Hb). Sickle cell disease (SCD) is an autosomal
recessive haemoglobinopathy resulting from a single nucleotide polymorphism (SNP)
leading to one nucleotide substitution (GTG for GAG) in the 6th codon of the bglobin gene. In sub-Saharan Africa, the Hb disorders are mainly associated to the
pathological variants hemoglobin S (HbS) and HbC. Most applicable diagnostic
techniques are based on amino acid detection and often require advanced
laboratory equipment. Therefore, it is challenging to acquire a DNA-based and
affordable diagnosis of SCD in sub-Saharan Africa. The aim of this study was to
develop a molecular assay that targets the point-mutation within HbS gene. Two
molecular assays were proposed; one primer-specific LAMP assay and one
Restriction Enzyme-RPA assay (RE-RPA). Both molecular reactions are isothermal and
widely used at the point-of-care. The specific LAMP assay was combined with a novel
portable device that allows real-time colorimetric LAMP experiments. The RE-RPA
assay was combined with the acoustic technology of Quartz Crystal Microbalance
with Dissipation. The first assay enabled the discrimination of the wild type genotype
(HbAA) and with the second assay the HbSS mutant genotype differentiation from
the HbAA was achieved. By furtherly optimizing the RE-RPA assay, we hope to
subsequently demonstrate a one-step efficient DNA amplification with high
reliability at the point-of-care.
|
Language |
English |
Subject |
Biosensor |
|
Isothermal reaction |
|
Βιοαισθητήρας |
|
Ισοθερμική αντίδραση |
Issue date |
2022-11-25 |
Collection
|
School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses
|
|
Type of Work--Post-graduate theses
|
Views |
410 |