DEVELOPMENT OF A PORTABLE, LOW-COST PLATFORM FOR NUCLEIC ACID AMPLIFICATION AND DETECTION

Abstract

Nucleic acid amplification tests (NAATs) play a critical role in clinical diagnostics by enabling rapid and highly sensitive detection of genomic sequences associated with specific pathogens. While fluorescence sensing is a common detection modality for NAATs, current fluorescence readers are large, expensive, or lack integrated thermal control. This thesis presents the development of a compact and low-cost assay reader combining precise thermal control and multiplexed fluorescence detection that can be adapted for a wide range of NAAT formats. The portable and wireless system, termed the Multiplexed Array Gene Imager (MAGI), has a volume of 380 cm3 and costs less than $90 to build. The MAGI system can be powered using only a battery pack and is controlled wirelessly by a computer using a Web-based graphical user interface. Performance of the MAGI system was evaluated using loop-mediated isothermal amplification assays that achieved time-to-positive values under 20 min. Overall, the MAGI system is easy to manufacture and assemble and provides a highly flexible and functional platform that can be adapted to microfluidic or microwell based assays for use in diverse environments including point-of-care and at-home settings.