A shortened 8-mer ssDNA aptamer was successfully truncated for four different tetracyclines with high affinity. The ultrasensitive colorimetric detection of oxytetracycline using this shortened aptamer was possible, which was about 500-fold enhanced compared to that obtained using the original 76-mer aptamer.
We describe a simple, high-speed, high-throughput aptamer screening for a group of small molecules using graphene oxide (simple Multi-GO-SELEX) without immobilizing targets. The affinities of ten different ssDNA aptamers successfully obtained for three pesticides were in the range of 10-100 nM. Besides a specific aptamer for each target, we found a couple of flexible multi-target aptamers, which can bind with 2 or 3 different molecules. These flexible aptamers developed for binding with a mixture of targets are not only significant for the rapid screening of a group of small molecules but also offer great promise for aptamer-based biosensor applications.
Porcine reproductive and respiratory syndrome virus (PRRSV) causes porcine reproductive and respiratory syndrome disease (PRRS), a disease that has a significant and economic impact on the swine industry. In this study, single-stranded DNA (ssDNA) aptamers with high specificity and affinity against VR-2332 strain of PRRSV type II were successfully obtained. Of 19 candidates, the LB32 aptamer was found to be the most specific and sensitive to VR-2332 strain according to an aptamer-based surface plasmon resonance (SPR) analysis. The detection of VR-2332 of PRRSV type II was successfully accomplished using the enzyme-linked antibody-aptamer sandwich (ELAAS) method. The detection limit of ELAAS was 4.8 × 10(0) TCID(50)/mL that is comparable to some of the previous reports of the PCR-based detection but does not require any complicated equipment or extra costs. Moreover, this ELAAS-based PRRSV detection showed similar sensitivity for both the VR-2332 samples spiked in diluted swine serum and in buffer. Therefore, this VR-2332 strain-specific aptamer and its assay method with high specificity can be used as an alternative method for the fast and precise detection of PRRSV.
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