Development of a microfluidic based system to detect Legionella pneumophila using Polymerase chain reaction

The objective of this doctoral dissertation is to develop a microfluidic-based system for the detection of Legionella using polymerase chain reaction (PCR). To achieve this objective, three principal components are pursued throughout the course of this study. The initial objective is to develop an automated system for sample preparation. This will be followed by the creation of a fluidic system for mixing and aliquoting the sample. Finally, a thermal cycler will be developed that is suitable for the process. It will be demonstrated that a concentration of samples is feasible with the assistance of a filtration and backwashing procedure. It will become evident that an automated configuration for this process is to be pursued in order to minimize the potential for human error. Additionally, a variety of mixing strategies were evaluated, and a conceptual framework for aliquoting was developed. The use of simulations enabled the evaluation and optimization of systems. The previously simulated system, comprising a mixer and a flow splitter, was produced using a subtractive process. Subsequently, the mixing and aliquoting capabilities of the system could be investigated in the laboratory assembly. Ultimately, a cost-effective PCR thermocycler was developed, wherein the distinct temperature parameters of a PCR run could be met with precision in a fundamental configuration. Three distinct design variants were evaluated for the basic structure.