Multiplexed lateral flow biosensors: Technological advances for radically improving point-of-care diagnoses

• Multiple analyte detection on paper-fluidic devices.
• Multi-parameter lateral flow detection.
• Spatial multiplexity by 2D paper network.
• Conceptual multiplexity by logic gate-based lateral flow biosensor.
• Sensitivity and specificity of multiplexed lateral flow detection.
• Future perspectives of multiplexed lateral flow biosensors.

Lateral flow biosensors are a leading technology in point-of-care diagnostics due to their simplicity, rapidness and low cost. Their primacy in this arena continues through technological breakthroughs such as multiplexing: the detection of more than one biomarker in a single assay. Multiplexing capacity is critical for improving diagnostic efficiency, enhancing the diagnostic precision for specific diseases and reducing diagnostic cost. Here we review, for the first time, the various types and strategies employed for creating multiplexed lateral flow biosensors. These are classified into four main categories in terms of specific application or multiplexing level, namely linear, parameter, spatial and conceptual. We describe the practical applications and implications for each approach and compare their advantages and disadvantages. Importantly, multiplexing is still subject to limitations of the traditional lateral flow biosensor, such as sensitivity and specificity. However, by pushing the limitations of the traditional medium into the multiplex arena, several technological breakthroughs are emerging with novel solutions that further expand the utility of lateral flow biosensing for point-of-care applications.