A novel label free long non-coding RNA electrochemical biosensor based on green l-cysteine electrodeposition and Au–Rh hollow nanospheres as tags

Abstract

Nuclear paraspeckle assembly transcript 1 (NEAT1) that reflects whether the body's anti-HIV virus immunity is strong or weak is one of many long non-coding RNAs (lncRNAs). The effective detection of NEAT1 has great scientific significance and clinical value. However, the detection of lncRNA is difficult with existing methods. The quantification of lncRNA by electrochemical methods based on changes in circuit properties such as capacitance, conductance and resistance may solve this problem. In this study, we describe a new label-free strategy based on catalytic signal amplification with single-wall carbon nanotubes wrapped with Au-Rh hollow nanospheres (Au/Rh-HNP@SWCNT). This strategy is useful for detecting the lncRNA NEAT1. Firstly, L-cysteine (L-Cys) was electrodeposited onto a Au electrode and incubated in colloidal Au to fully expose all of the binding sites of the Au particles, which can bind to L-Cys through its thiol-group. It can take advantage of each surface of the Au nanoparticles that is bound to the capture probe, which contains a (GGG) 3 trimer that can bind to the electron mediator hemin. The results indicate that catalysis was noticeably enhanced and that the biosensor provided ultrasensitive detection of the lncRNA NEAT1. The linear calibration of the biosensor ranged from 1 fM mL À1 to 100 nM mL À1 , and the limit of detection was 0.8863 fM mL À1 . This lncRNA biosensor based on Au/Rh-HNP@SWCNT complex signal amplification and an L-Cys Au nano-film exhibited acceptable reproducibility and clear selectivity. This strategy may provide a new alternative for clinical HIV diagnosis through the detection of NEAT1.

RNA Biomarkers: Diagnostic and Prognostic Potentials and Recent Developments of Electrochemical Biosensors

“…Moreover, a simplified electrochemical sensor using direct interaction of target RNA and gold electrode has also been developed to circumvent the multisteps sensor fabrication steps . We have also observed the recent trends of using novel nanomaterials to accelerate the signal transduction and obtain amplified and sensitive detection signal . Despite the rapid advances in the biosensing of mRNAs and circulating miRNAs, development of electrochemical sensor for lncRNAs detection is still in its infancy.…”

“…Despite the huge clinical significance of lncRNAs, their electrochemical detection is not well explored. Until now, only a few reports on electrochemical biosensing of lncRNAs are available . This could be explained by the fact that lncRNA has a relatively long sequence with a high molecular weight.…”

“…Taken together these issues, the commonly used RNA‐hybridization‐based electrochemical sensors cannot recognize and effectively capture the target lncRNAs on the electrode surface. In 2015, Liu et al used the catalytic amplification ability of single‐wall carbon nanotubes coated with Au–Rh hollow nanospheres (Au/Rh‐HNP@SWCNT) to detect nuclear paraspeckle assembly transcript 1 (NEAT1) lncRNA, which is reportedly overexpressed in patients with HIV. The nanospheres have the combined benefits of RNA binding features of gold, unique electronic properties, and large edge‐plane‐to‐basal‐plane ratio of single‐wall carbon nanotubes (SWCNTs), and the surpassing catalytic properties of Rh.…”

Nanomaterial-based biosensors for detection of pathogenic virus

“…In the case of virus detection, Pu et al [48] used the ability of surface decorating of CNTs for making them ready to immobilize on gold sheets to synthesize a modified electrode. Horseradish peroxidase enzyme was set on carbon nanotubes which contain capture probes, and then joined to the gold layers by Au nanoparticles.…”

PCR-Free Detection of Long Non-Coding HOTAIR RNA in Ovarian Cancer Cell Lines and Plasma Samples

“…Additionally, these assays provide intrinsic simplicity, portability, and a high potential for miniaturized, multiplexed, and decentralized analysis of microRNA and other nucleic acid biomarkers with an elevated translational capacity [22,23]. Despite these developments, very few examples of lncRNA biosensors have been reported to date [26,27]. A biosensor with dual readout ability that couples naked-eye visualisation (colorimtric signal) with electrochemical readout is highly compatible for biomolecular sensing, particularly in poor resource environments, where naked-eye evaluation could be used as the first screening pass of the analyte.…”