Lateral flow assays for Ochratoxin A using metal nanoparticles: comparison of “adsorption–desorption” approach to linkage inversion assembled nano-aptasensors (LIANA)

Abstract

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.