<p>Enteroviruses
are a ubiquitous mammalian pathogen that can produce mild to life-threatening
disease. Bearing this in mind, we have developed a rapid, accurate and
economical point-of-care biosensor that can detect a nucleic acid sequences
conserved amongst 96% of all known enteroviruses. The biosensor harnesses the
physicochemical properties of gold nanoparticles and aptamers to provide
colourimetric, spectroscopic and lateral flow-based identification of an
exclusive enteroviral RNA sequence (23 bases), which was identified through in
silico screening. Aptamers were designed to demonstrate specific
complementarity towards the target enteroviral RNA to produce aggregated
gold-aptamer nanoconstructs. Conserved target enteroviral nucleic acid sequence
(≥ 1x10<sup>-7</sup> M, ≥1.4×10<sup>-14</sup> g/mL), initiates gold-aptamer-nanoconstructs
disaggregation and a signal transduction mechanism, producing a colourimetric
and spectroscopic blueshift (544 nm (purple) > 524 nm (red)). Furthermore,
lateral-flow-assays that utilise gold-aptamer-nanoconstructs were unaffected by
contaminating human genomic DNA, demonstrated rapid detection of conserved target
enteroviral nucleic acid sequence (< 60 s) and could be interpreted with a
bespoke software and hardware electronic interface. We anticipate our
methodology will translate in-silico screening of nucleic acid databases to a
tangible enteroviral desktop detector, which could be readily translated to
related organisms. This will pave-the-way forward in the clinical evaluation of
disease and complement existing strategies at overcoming antimicrobial
resistance.</p>
Construction Strategy for an Internal Amplification Control for Real-Time Diagnostic Assays Using Nucleic Acid Sequence-Based Amplification: Development and Clinical Application