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

• Autores: Olena Mayboroda
• Directores de la Tesis: Ioanis Katakis (dir. tes.), Ciara K. O'Sullivan (codir. tes.)
• Lectura: En la Universitat Rovira i Virgili ( España ) en 2017
• Idioma: español
• Tribunal Calificador de la Tesis: Mamantos Prodromidis (presid.), Mayreli Ortiz Rodríguez (secret.), Luis Antonio Tortajada-Genaro (voc.)
• Programa de doctorado: Programa de Doctorado en Nanociencia, Materiales e Ingeniería Química por la Universidad Rovira i Virgili
• Materias:
  • Química
    • Química analítica
    • Bioquímica
  • Ciencias de la vida
    • Biología molecular
• Enlaces
  • Tesis en acceso abierto en: TDX
• Resumen

  • An easy, rapid and reliable detection of multiple biological warfare agents, pathogens, viruses and toxins is required in different situations of molecular diagnostics: from personalised medicine to bioterrorism defence. The main goal of this Doctoral thesis is to present alternative amplification and detection platform that combines the isothermal amplification technique with a solid-phase detection system in order to overcome some of the currently existing limitations in the field.

    This work presents a simple, rapid and easy to integrate isothermal amplification protocol that can be used for analysis of genetic material and has good chances to be applied in point of care devices. Ultimately, the proved multiplexing ability of the system makes it a promising candidate to be integrated in devises for in situ testing due to its simplicity and reasonable pricing.

    The thesis has the following structure: Chapter 1 is an introduction to the topic with description of the current trends and problems in the field of molecular diagnostics, the role of isothermal DNA amplification and biosensors in evolution of the field, the current state of the art and the objectives of this thesis. Chapters 2-5 report the work performed in order to achieve the specific objectives of this Doctoral thesis: the development of homogeneous multiplex isothermal system for detection of several pathogens in one pot, the use of solid-phase recombinase polymerase amplification strategy using optical and electrochemical detection, the exploiting of the surface chemistry to overcome the limitations found, and the use of labelled dNTPs in the amplification process in order to simplify and shorten the time of the assay. The conclusions and the future prospects are described at the end of the thesis.