Abstract
This study aimed to prepare two hairpin-structure DNA probes by conjugating carminic acid (CA) or hemin into two ends of specific genes of coxsackievirus A16 (CV-A16) and enterovirus A71 (EV-A71) (probeCV-A16-CA and probeEV-A71-hemin). Then, probeCV-A16-CA and probeEV-A71-hemin as the signal molecules were adsorbed onto NH2-MIL-53 (Al) (MOF). Based on these biocomposites, an electrochemical biosensor with dual-signal outputs for simultaneous assay of CV-A16 and EV-A71 was constructed. The stem-loops of probes switched both CA and hemin monomer to dimer, reducing the electrical activity of both CA and hemin. Subsequently, the target-induced opening of the stem-loop switched both CA and hemin dimers to monomers, resulting in two nonoverlapping increasing electrical signals. This sensitively reflected the concentration of targetCV-A16 and targetEV-A17 ranging from 10−10 to 10−15 M with a detection limit of 0.19 and 0.24 fM. This strategy was mainly applied to the simultaneous determination of targetCV-A16 and targetEV-A17 in 100% serum with satisfactory results. The MOF combined with the high loading capacity broke through the intrinsic limitation on sensitivity using the traditional methods. An increase of three orders of magnitude was observed. This study involved simple one-step detection, and only a simple replacement of a gene could trigger its potential in clinical and diagnostic applications.
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