Development of diagnostic platform for detection of biological agents and toxic microalgae using isothermal amplification

Identificador:  TDX:2720

Handle: https://hdl.handle.net/20.500.11797/TDX2720

Autors:  Mayboroda, Olena

Resum:
The main goal of this doctoral thesis is to present alternative approaches in the field of DNA biosensors, design and develop new isothermal amplification protocols compatible with a portable, easy-to use device that can be deployed for analysis of genetic material at the point-of-care/need, while overcoming some of the currently existing limitations. In order to achieve this objective, a variety of different strategies of effective solid-phase immobilisation and liquid-phase and solid-phase isothermal enzymatic amplification have been studied to achieve lower detection limits with rapid and easy to carry out assays. This work presents a convenient, flexible solution for detecting DNA with biosensors, exploiting a general concept of liquid-phase and solid-phase isothermal amplification and detection, thus integrating two nucleic acid tests, PCR and microarrays, in one single device. The thesis report the work performed to achieve the specific objectives of this doctoral thesis: the development of multiplexing platform for simultaneous detection of several targets, the use of solid-phase recombinase polymerase amplification strategy for DNA amplification with optical and electrochemical detection, the detection of real samples, the study of the surface chemistry and the combination of isothermal amplification with redox labelled dNTPs for the amplification and detection of toxic microalgae as an innovative method that permits incorporation of labels throughout the amplification process facilitating direct electrochemical detection of the DNA products and an inherent shortening of assay time.