Methods for the formation of gold-aptamer nanoparticle constructs were adapted from Liu and Lu. Briefly, Sequence-A (9 μL, 1 mM) and a mixture of Sequence-A (4.5 μL, 1 mM) & Sequence-B-Biotin (4.5 μL, 1mM) were added to two separate sodium hydroxide (12 M) cleaned vials. Tris(2-carboxyethyl) phosphine (TCEP, 1.5 μL, 10 mM) and acetate buffer (pH 5.2, 1μL, 500 mM) were added to each vial and incubated at room temperature for 1 hr. Gold nanoparticle suspensions (3 mL) were added to each vial and stirred overnight. Tris acetate buffer (pH 8.2, 30 μL 500 mM) and sodium chloride solution (300 μL, 1 M) were added dropwise to each vial and stirred overnight. Functionalised gold nanoparticle suspensions were transferred to micro-centrifuge tubes and washed with sodium chloride (100 mM)/ Tris acetate (25 mM) solution and centrifugation (1 mL, 3 times, 16,110 g, 15 min) discarding the supernatant after each wash. After the final wash the content of the vials containing gold nanoparticles conjugated to Sequence-A and a mixture of Sequence-B and Sequence-B-Biotin were combined in a single vial suspended in sodium chloride (300 mM)/ Tris acetate (25 mM) buffer solution (1 mL). To this vial the aptamer strand was added (60 μL, 10 μM) to hybridise with aptamer functionalised gold and could aggregate overnight under refrigeration (4 °C). The following day the aggregated nanoparticles were washed with sodium chloride (600 mM)/ Tris acetate (25 mM) buffer solution and centrifugation (1 mL, 3 times, 800 g, 2 min) to eliminate free sensory aptamer. When the detection limit is 1x10 M and the ratio of target conserved enteroviral nucleic acid to aptamer to Sequence-A, Sequence-B and Sequence-B-Biotin is 1:1:0.5:0.5, in a 1 mL sample of concentrated gold-aptamernanoconstructs, the amount of nucleic acid bound to gold nanoparticle surfaces is estimated to be 0.1 nmol for Sequence-A and 0.05 nmol for Sequence-B and Sequence-B-Biotin. The final pellet was suspended in sodium chloride (600 mM)/ Tris acetate (25 mM) buffer solution (3 mL) and was ready to use for experimentation or stored under refrigeration (4 °C).