Detection of low abundance target DNA/RNA for clinical or research purposes is challenging because the target sequences can be hidden under a large background of human genomic or non-human metagenomic sequences. We describe a probe-based capture method to enrich for target sequences with DNA-clicked iron oxide nanoparticles. Our method was tested against commercial capture assays using streptavidin beads, on a set of probes derived from a common genotype of the hepatitis C virus.
Fig: Description of InBeads capture system. (a) DNA-conjugated IONPs are incubated with a target solution to promote hybridization. Non-specifically hybridized DNA/RNA are removed by washes. Specifically hybridized DNA/RNA are retained on the magnetically immobilized beads. (b) Preparation of InBeads. (i) IONPs synthesis, (ii) silica coating, (iii) azide functionalization, and (iv) conjugation with DNA probes through click chemistry.
We showed that our method is more specific and sensitive, most likely due to the combination of an inert silica coating and a high density of DNA probes clicked to the nanoparticles. This facilitates target capture below the limits of detection for TaqMan qPCR, and we believe that this method has the potential to transform management of infectious diseases.
Damavandi, F., Wang, W., Shen, WZ. et al. Enrichment of low abundance DNA/RNA by oligonucleotide-clicked iron oxide nanoparticles. Sci Rep 11, 13053 (2021). https://doi.org/10.1038/s41598-021-92376-9