- Title
- Development of aptamers against epitopes of the Ebola virus nucleoprotein for future applications in diagnostics
- Creator
- Mutombwera , Atherton Tiripano
- Subject
- Ebola virus disease Ebola virus disease -- Treatment
- Subject
- Epidemics
- Date Issued
- 2016
- Date
- 2016
- Type
- Thesis
- Type
- Masters
- Type
- MSc
- Identifier
- http://hdl.handle.net/10948/45931
- Identifier
- vital:39321
- Description
- Five different subtypes of the Ebola virus (EBOV) have been described. Either Zaire or Sudan EBOV subtypes has caused all of the EBOV outbreaks to date. The March 2014 Zaire EBOV disease outbreak that ravaged West Africa had a mortality rate of 70%, and resulted in 11 315 deaths. Swift cost effective EBOV detection is required to manage EBOV disease outbreaks as this leads to the interruption of the chain of transmission. Lateral flow diagnostic devices (LFDs) have been designed to provide quick, simple and cost effective diagnosis at the point of care and have great potential at interrupting the chain of EBOV transmission. The target recognition elements used in LFDs are the most important components of an LFD as they determine not only the selectivity and specificity of the device but also the transportation and storage conditions of the devices. Antibodies are the most common biomolecules used as target recognition elements in LFDs. However, the cost of producing antibodies is high and these biomolecules are highly sensitive to changes in the environmental conditions (e.g. temperature, pH and ionic strengths of buffer conditions), which can affect the selectivity and specificity of the LFDs. Aptamers can be used as alternative target recognition elements in LFDs. Aptamers are short single stranded nucleic acid molecules that have the ability to bind to their targets (e.g. whole cells, small molecules, toxins, proteins and peptides) with high affinity and specificity. By replacing antibodies with aptamers, LFDs can be produced that are less expensive, have higher selectivity and specificity. The aim of this study was to generate aptamers against the two highly conserved linear epitope regions (amino acid 421-440 and amino acid 601-620) of the EBOV nuclear protein (NP) using site directed SELEX. Such aptamers can be used as target recognition elements in the development of a LFD for the diagnosis of EBOV infection. Four aptamers that can potentially bind to the linear epitope spanning from amino acid 421 to 440 of the EBOV NP and four aptamers that can potentially bind to the linear epitope spanning from amino acid 601 to 620 of the EBOV NP were identified in this study. An in silico analysis of the predicted secondary structure of the putative aptamers was performed before and after the truncation of nucleotide sequences from the 5’ and 3’ ends of the aptamers to remove excess nucleotide sequences. Although this study did not characterise the interaction between the aptamers and linear epitope regions, the study succeeded in optimising the buffer conditions for future interaction studies using the SPR Biacore 3000 instrument.
- Format
- x, 94 leaves
- Format
- Publisher
- Nelson Mandela Metropolitan University
- Publisher
- Faculty of Science
- Language
- English
- Rights
- Nelson Mandela Metropolitan University
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NMMU School of BioMolecular and Chemical Sciences
| Thumbnail | File | Description | Size | Format | |||
|---|---|---|---|---|---|---|---|
| View Details Download | SOURCE1 | Atherton Tiripano Mutombwera.pdf | 2 MB | Adobe Acrobat PDF | View Details Download |