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Thallium(i) and lead(ii) ions are heavy metals and extremely toxic. These metals are environmental pollutants, posing a severe risk to the environment and human health. In this study, two approaches were examined using aptamer and nanomaterial-based conjugates for thallium and lead detection. The first approach utilized an in-solution adsorption-desorption approach to develop colorimetric aptasensors for the detection of thallium(i) and lead(ii) using gold or silver nanoparticles. The second approach was the development of lateral flow assays, and their performance was tested with thallium (limit of detection is 7.4 μM) and lead ion (limit of detection is 6.6 nM) spiked into real samples. The approaches assessed are rapid, inexpensive, and time efficient with the potential to become the basis for future biosensor devices., Competing Interests: The authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)

In this study, a FRET-based fluorescent aptasensor for the detection of ochratoxin A (OTA) was optimized based on the quenching efficiency of single-walled carbon nanotubes (SWCNTs) and the binding affinity of aptamers. OTA aptamers were conjugated with quantum dots and adsorbed to the surface of both acid-modified and unmodified SWCNTs. The maximum fluorescence quenching efficiency of the SWCNTs were compared. Acid-modified SWCNTs (amSWCNTs) have moderate quenching efficiency, providing an optimal sensitivity for qualitative fluorescence-enhancement biosensor assays. The binding parameters of the QD-modified OTA aptamers (1.12.2 and A08min) on the surface of amSWCNTs were compared. Based on our results, the A08min aptamer is a better candidate for OTA detection. Using the A08min aptamer, the SWCNT method had a limit of detection (LOD) of 40 nM. The amSWCNT method had a significantly lower LOD of 14 nM. Turn-on fluorescent nano-aptasensors are emerging as an effective diagnostic tool for simple detection of mycotoxins. Nanocomplexes designed for the detection of mycotoxins in solution and paper-based tests have proven to be useful., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Targeting of molecular constituents of thrombi with aptamer functionalized core-shell nanoparticles (CSN) allowed for high resolution clot delineation in T2-weighted magnetic resonance imaging. Meanwhile, the gold-coating demonstrated sufficient contrast capabilities in computed tomography (1697 HU μM -1 ). This targeted CSN formulation could allow for precise imaging of blood clots at low nanomolar concentrations.

Nano-aptamer probes were prepared and used in lateral flow colorimetric assays for the detection of Ochratoxin A (OTA). In this study, two approaches were examined using 5'-biotin-modified OTA aptamers and silver or gold nanoparticles (AgNP or AuNP). The first method used an "adsorption-desorption" approach wherein aptamers were adsorbed onto the metal nanoparticle surface. Upon the addition of OTA, the aptamer binds specifically to the target, releasing the NPs. The above solutions were applied on a lateral flow assay (LFA) and a detection limit of 6.3 nM was achieved with both metal nanoparticles. The second method used a labelled approach based on Linkage Inversion Assembled Nano-Aptasensors (LIANAs) using a DNA linker containing a 5'-5' linkage inversion (5'-5' linker) to assemble biotinylated aptamer-functionalized metal nanoparticles. In the presence of target, OTA specifically binds with its aptamer leading to release of the linker and disassembly of LIANA aggregates into dispersed nanoparticles. The same solutions were applied in LFA format and the lowest detection limit of 0.63 nM was achieved. The results indicated that the LIANA-based LFA strips were more sensitive than the "adsoprtion-desorption" LFAs. Both lateral flow assays are inexpensive, simple, and rapid to perform and produces results visible to the naked-eye.

Aptamers are short single-stranded oligonucleotides that fold into unique three-dimensional structures, facilitating selective and high affinity binding to their cognate targets. It is not well understood how aptamer-target interactions affect regions of structure in an aptamer, particularly for small molecule targets where binding is often not accompanied by a dramatic change in structure. The DNase I footprinting assay is a classical molecular biology technique for studying DNA-protein interactions. The simplest application of this method permits identification of protein binding where DNase I digestion is inhibited. Here, we describe a novel variation of the classical DNase I assay to study aptamer-small molecule interactions. Given that DNase I preferentially cleaves duplex DNA over single-stranded DNA, we are able to identify regions of aptamer structure that are affected by small molecule target binding. Importantly, our method allows us to quantify these subtle effects, providing an in solution measurement of aptamer-target affinity. We applied this method to study aptamers that bind to the mycotoxin fumonisin B1, allowing the first identification of high affinity putative minimers for this important food contaminant. We confirmed the binding affinity of these minimers using a magnetic bead binding assay.

A strategy for aptamer-based biosensing termed linkage inversion assembled nano-aptasensors (LIANAs) is shown to be a generally applicable approach to the sensitive and specific detection of a target molecule in turn-on fluorescence solution-based and paper-based tests.

A new approach to the development of pH-driven DNA switches is presented. Here, the improved stability of the A x G mispair under slightly acidic conditions is exploited. The temperature tunability and the potential generality of the approach are discussed.