Strategies for the Development of Multiplexed Diagnostics

Abstract

Lateral flow assays (LFAs) are the most common form of a rapid diagnostic test, and their use has become critical for public health surveillance. These immunoassays allow for quick time-to-result and allow for users to self-operate the device and interpret their own results. Although LFAs are widely used, they are developed empirically, leaving substantial room for analysis and optimization. Multiplexed diagnostics are crucial tools for the simultaneous detection of multiple analytes and their use has become more abundant in advanced diagnostic techniques, such as polymerase chain reaction. These diagnostics have increased utility when compared to single-plexed diagnostic tools and there is a consensus that more of these diagnostics are needed, specifically multiplexed diagnostics that can be used at the point-of-care. Although the utility of multiplexed diagnostics is well-recognized, the development and commercialization of these tools, especially when conducted in an LFA format, is hindered by several obstacles. The most prominent of these obstacles is cross-reactivity. This work uses inductively coupled optical emission spectroscopy to analyze gold distribution on commercially available LFAs, identifying key areas of improvement. Various methods for the development of an in-house multiplexed LFA are discussed also along with strategies to combat inherent cross-reactivity issues.