The spectroscopic (colour and ultraviolet UV absorbance maxima) and physicochemical properties (mobility across lateral flow membranes) of gold nanoparticles differ when they are in disaggregated or aggregated constructs. Therefore, prior to harnessing these properties to develop a biosensor to detect conserved target enteroviral nucleic acid sequences, gold nanoparticles and gold-aptamer-nanoconstructs were synthesised, functionalised with Sequence-A, Sequence-B and aptamer, as well as characterised for their spectroscopic properties, size and zeta potential.