Novel Analytical Systems for Rapid Forensic Nucleic Acid Detection

Forensic DNA analysis is a discipline in criminalistics founded by the pillars of molecular biology and analytical techniques, whereby the generation of a DNA profile is used for human identification (hID). In recent years, increased awareness for backlogged sexual assault kits and demand in automated processing of reference samples have prompted changes in the workflow of hID. Improvements to the assay accuracy, efficiency, and automation are attainable due to the advent of novel analytical methods and continued innovation in technology.
An introduction to the processes involved in hID is provided in Chapter 1, as well as the relevant literature surrounding the conventional and emerging methods in screening sexual assault evidence. Additionally, the current status and technical maturity of micro total analysis systems in both commercial and academic sectors are presented. The aim of the work presented in Chapter 2 and 3 was to develop a novel method for screening sexual assault evidence, with the intent of addressing the testing bottleneck currently experienced in forensic laboratories. The optimization of this ‘Y-screening’ assay was detailed in Chapter 2, involving an enzymatic lysis protocol and the subsequent colorimetric loop-mediated isothermal amplification method. A thorough assessment of the assay’s reliability, capability, and limitations was characterized in Chapter 3, as well as the documentation of the development of a prototype integrated system for performing high-throughput, objective Y-screening assays. Chapter 4 and 5 present the work conducted towards the completion of the faSTR project – a multi-year effort to develop a new generation of portable, automated, and integrated system for hID applications. Chapter 4 describes the optimization of an automated method to align the optics to a 50-micron electrophoresis channel, based on innate fluorescent signals. The final evaluation of the faSTR system, both in terms of hardware and assay performance, is presented in Chapter 5.