Abstract
MiRNA-150, a gene regulator that has been revealed to be abnormal expression in non-small cell lung cancer (NSCLC), can be regarded as a serum indicator for diagnosis and monitoring of NSCLC. Herein, a new sort of nanoprobe, termed allosteric spherical nanoprobe, was first developed to sense miRNA-150. Compared with conventional hairpin, this new nanoprobe possesses more enrichment capacity and reaction cross section. Structurally, it consists of magnetic nanoparticles and dual-hairpin. In the absence of miRNA-150, the spherical nanoprobes form hairpin structure through DNA self-assembly, which could promote the Förster resonance energy transfer (FRET) of fluorophore (FAM) and quencher (BHQ1) nearby. However, in the presence of target, the target-probe hybridization can open the hairpin and form the active “Y” structure which separated fluorophore and quencher to yield “signal on” fluorescence. In the manner of multipoint fluorescence detection, the target-bound allosteric spherical nanoprobe could provide high detection sensitivity with a linear range of 100 fM to 10 nM and a detection limit of 38 fM. More importantly, the proposed method can distinguish the expression of serum miRNA-150 among NSCLC patients and healthy people. Finally, we hoped that the potential bioanalytical application of this nanoprobe strategy will pave the way for point-of-care testing (POCT).
Highlights
MicroRNAs are sorts of small single stranded noncoding RNAs and its length are approximately 21 ~ 25 nucleotides [1,2,3]
To upgrade the assay sensitivity, most of these determinations were carried out assist with signal amplification strategies, including rolling circle amplification (RCA), exponential amplification reaction (EXPAR), catalytic hairpin assembly (CHA), entropy driven catalytic reaction, enzyme-assisted signal amplification and materials, etc. [21,22,23,24,25,26]
Design principle of allosteric spherical nanoprobe As shown in Scheme 1, the allosteric spherical nanoprobe was structurally consisted of three sequences (Seq A: the free chain, Seq B: the fixed chain, Seq C: the auxiliary chain) and formed double rings that were hybridized with target nucleic acids
Summary
MicroRNAs (miRNAs) are sorts of small single stranded noncoding RNAs and its length are approximately 21 ~ 25 nucleotides [1,2,3]. Since the dysregulation of miRNAs is closely related to the occurrence and prognosis of tumor, the detection of circulating miRNAs in serum can be Currently, the detection of miRNAs is normally performed by traditional techniques such as northern blotting, microarray and reverse transcription-polymerase chain reaction (RT-PCR) [7,8,9]. These methods are reliable, they are hardly applied to clinic due to. It is expected to establish one kind of simpler method for target directly detection without the help of any amplification strategy
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