A microwave powered DNA extraction module

Wilson-Garner, Solstice 2023. A microwave powered DNA extraction module. PhD Thesis, Cardiff University. Item availability restricted.

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Abstract

Mycobacterium tuberculosis is a multidrug resistant pathogen commonly isolated from patients with TB. Currently, there is no rapid diagnostic tool to detect the presence of M.tuberculosis. The rapid diagnosis and prompt subsequent treatment remains the best curative approach to reduce disease burden and interrupt transmission. A major concern in developing a rapid diagnostic assay is DNA extraction. Mycobacterial cells are very difficult to lyse; the existing methods are time consuming, resulting in long process time to detect the pathogen. The aim of this project was to improve upon an existing method of DNA extraction and verification by creating a 10ml cartridge capable of promoting microwave interaction with entire TB patient samples (as opposed to 250μl samples used in the past) in a single excitation stage, thereby vastly improving the sensitivity of the assay reducing the risk of false negative results. The microwave applicator parameters were optimised to obtain the maximum possible DNA release from the target pathogen. This DNA was then collected by means of DNAspecific probes attached to magnetic nanoparticles, allowing the target DNA to be separated and concentrated before it was detected with a lower limit of 104CFU/ml. Finally, the project explored the possibility of integration of each stage of the diagnostic process (microwave application, magnetic nanoparticle collection, diagnosis) into a single portable device. In conclusion, a 10ml cartridge has been designed and microwave application parameters have been optimised to yield a rapid diagnostic assay. Initial exploration is underway to transfer this assay from the laboratory to real-world application.

Item Type:	Thesis (PhD)
Date Type:	Completion
Status:	Unpublished
Schools:	Schools > Pharmacy
Subjects:	Q Science > Q Science (General)
Date of First Compliant Deposit:	20 March 2025
Last Modified:	20 Mar 2025 15:14
URI:	https://orca.cardiff.ac.uk/id/eprint/176992

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