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Track-etched membranes as a platform for digital loop-mediated isothermal amplification
Thesis (MEng)--Stellenbosch University, 2025.
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| Format: | Thesis |
| Language: | en_ZA |
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Stellenbosch : Stellenbosch University
2025
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| _version_ | 1867614037995421696 |
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| access_status_str | Open Access |
| author | Blignaut, Lanique |
| author2 | Perold, Willem J. |
| author_browse | Blignaut, Lanique Perold, Willem J. |
| author_facet | Perold, Willem J. Blignaut, Lanique |
| author_sort | Blignaut, Lanique |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Thesis (MEng)--Stellenbosch University, 2025. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/132074 |
| institution | Stellenbosch University (South Africa) |
| language | en_ZA |
| last_indexed | 2026-06-10T12:45:40.774Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/132074 Track-etched membranes as a platform for digital loop-mediated isothermal amplification Blignaut, Lanique Perold, Willem J. Engelbrecht, Anna-Mart Stellenbosch University. Faculty of Engineering. Dept. of Electrical and Electronic Engineering. Gene amplification Membranes (Biology) -- Thermal properties Bioinformatics UCTD Thesis (MEng)--Stellenbosch University, 2025. Blignaut, L. 2025. Track-etched membranes as a platform for digital loop-lediated isothermal amplification. Unpublished masters thesis. Stellenbosch: Stellenbosch University [online]. Available: https://scholar.sun.ac.za/items/c1c4df4a-3072-4c39-bdaa-f276db78eb8b ENGLISH ABSTRACT: Digital genetic detection platforms enable the separation and detection of small sample quantities with high sensitivity, specificity, and reproducibility. Traditional digital polymerase chain reaction (dPCR), however, requires complex instrumentation for thermocycling, microfluidic chip fabrication, and fluidic control. Here, digital loop-mediated isothermal amplification (dLAMP) on a commercial track-etched membrane (TM) platform was demonstrated as a simplified, isothermal alternative. A LAMP assay was developed and evaluated on a real-time PCR machine, and TM surfaces were characterised through scanning electron microscopy, atomic force microscopy, and contact angle measurements. To create dLAMP droplets on the TM, a simple polydimethylsiloxane (PDMS) peel-off method was performed and detected through fluorescent microscopy. Capillary forces enabled liquid insertion into the pores and the excess LAMP assay was successfully removed from the TM surface. Isolated compartments, where positive and negative pores could be clearly distinguished, was achieved. Therefore, TMs were identified as suitable platforms for dLAMP. This simple, inexpensive, and rapid detection method is recommended for the absolute quantification of target genes.sample quantities with high sensitivity, specificity, and reproducibility. Traditional digital polymerase chain reaction (dPCR), however, requires complex instrumentation for thermocycling, microfluidic chip fabrication, and fluidic control. Here, digital loop-mediated isothermal amplification (dLAMP) on a commercial track-etched membrane (TM) platform was demonstrated as a simplified, isothermal alternative. A LAMP assay was developed and evaluated on a real-time PCR machine, and TM surfaces were characterised through scanning electron microscopy, atomic force microscopy, and contact angle measurements. To create dLAMP droplets on the TM, a simple polydimethylsiloxane (PDMS) peel-off method was performed and detected through fluorescent microscopy. Capillary forces enabled liquid insertion into the pores and the excess LAMP assay was successfully removed from the TM surface. Isolated compartments, where positive and negative pores could be clearly distinguished, was achieved. Therefore, TMs were identified as suitable platforms for dLAMP. This simple, inexpensive, and rapid detection method is recommended for the absolute quantification of target genes. AFRIKAANSE OPSOMMING: Digitale genetiese deteksieplatforms verskaf die moontlikheid vir skeiding en deteksie van baie klein monsterhoeveelhede met hoë sensitiwiteit, spesifisiteit en herhaabaarheid. Traditionele digitale polimerase kettingreaksie (dPKR) benodig egter komplekse instrumente vir die termo-siklusse, mikrovloeistof skyfie vervaardiging en vloeistofbeheer. Hier was digitale lus-gemedieerde isotermiese amplifikasie (dLAMP) op ’n kommersiële spoor-geëtste membraan (SM) platform gedemonstreer as ’n vereenvoudigde, isotermiese alternatief. ’n LAMP eksperiment is ontwikkel en getoets op ’n werklike-tyd PKR masjien en SM oppervlaktes was gekarakteriseer deur skandeer-elektron mikroskopie, atoomkrag mikroskopie en kontakhoek metings. Om dLAMP druppels op ’n SM te vorm, was ’n eenvoudige polimetielsiloksaan (PDMS) aftrek metode uitgevoer wat met fluoresserende mikroskopie waargeneem is. Kapillêre kragte het die inlating van die vloeistowwe in die porieë toeglaat, die oormaat vloeistowwe was suksesvol verwyder deur die aftrek tegniek en geisoleerde kompartemente, waar positiewe en negatiewe porieë duidelik geonderskei kan word, was verkry. SMe was dus geïdentifiseer as ’n geskikte platform vir dLAMP. Hierdie eenvoudige, relatiewe goedkoop en vinnige deteksie metode word aanbeveel vir die absolute kwantifisering van teikengene. Masters 2025-05-22T09:15:33Z 2025-05-22T09:15:33Z 2025-03 Thesis https://scholar.sun.ac.za/handle/10019.1/132074 en_ZA Stellenbosch University xvi, 119 pages : illustrations, includes annexures application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Gene amplification Membranes (Biology) -- Thermal properties Bioinformatics UCTD Blignaut, Lanique Track-etched membranes as a platform for digital loop-mediated isothermal amplification |
| title | Track-etched membranes as a platform for digital loop-mediated isothermal amplification |
| title_full | Track-etched membranes as a platform for digital loop-mediated isothermal amplification |
| title_fullStr | Track-etched membranes as a platform for digital loop-mediated isothermal amplification |
| title_full_unstemmed | Track-etched membranes as a platform for digital loop-mediated isothermal amplification |
| title_short | Track-etched membranes as a platform for digital loop-mediated isothermal amplification |
| title_sort | track etched membranes as a platform for digital loop mediated isothermal amplification |
| topic | Gene amplification Membranes (Biology) -- Thermal properties Bioinformatics UCTD |
| url | https://scholar.sun.ac.za/handle/10019.1/132074 |
| work_keys_str_mv | AT blignautlanique tracketchedmembranesasaplatformfordigitalloopmediatedisothermalamplification |