Accurate and Efficient Lipoprotein Detection Based on the HCR–DNAzyme Platform

Abstract

Cardiovascular disease is one of the main causes of death in the world, which is closely associated with dyslipidemia. Dyslipidaemia is usually manifested as a relatively higher level of low-density lipoprotein (LDL) and lower level of high-density lipoprotein (HDL). Thus, the quantitative detection of the LDL and HDL particles is of great importance to predict the risk of cardiovascular diseases. However, the traditional methods can only indirectly reflect the HDL/LDL particle concentrations by detecting the cholesterol or proteins in HDL/LDL particles and are always laborious and time-consuming. Thus, the accurate and efficient approach for the detection of intact HDL and LDL particles is still lacking so far. We developed an enzyme- and isolation-free method to measure the concentration of HDL and LDL based on DNAzyme and hybridization chain reaction (HCR)-based signal amplification. This method can be used to directly and accurately detect the concentration of “actual” HDL and LDL particles instead of the cholesterol in HDL and LDL, with limits of detection of 10 and 30 mg/dL, respectively, which also satisfied the lipoprotein analysis in clinical samples. Therefore, this HCR–DNAzyme platform has great potential in clinical applications and health management.

Recent Advances on DNAzyme‐Based Sensing

“…The Willner group has reported a series of nucleic acid based constitutional dynamic network to mimic natural processes by chemical means where DNAzymes act as signal reporter units [70] . In addition, through the integration of DNAzymes with non‐enzymatic DNA amplification circuits, including catalytic hairpin assembly (CHA), [71] hybridization chain reaction (HCR), [72] entropy‐driven circuits (EDC) [73] and other enzyme‐mediated amplification techniques, [74] the sensitivity of DNAzyme‐based molecular probes was greatly enhanced to achieve amplified nucleic acids detection in vitro and in live cells. For instance, the Wang group reported a novel isothermal nucleic acid amplification strategy that was designed to use CHA and HCR to construct DNAzyme amplifier, allowing the accurate detection of microRNA with a limit of detection of 1.…”

Ultrasensitive Homogeneous Detection of PCSK9 and Efficacy Monitoring of the PCSK9 Inhibitor Based on Proximity Hybridization-Dependent Chemiluminescence Imaging Immunoassay

“…Herein, a CL imaging approach was proposed for a wash-free assay of PCSK9 protein for the first time by integrating dual-recognition proximity hybridization, stimuli-responsive MSN, and glow-type CL reporter. Compared to most of the aptasensors based on single molecule recognition, the proposed proximity hybridization-dependent CL immunoassay (PCLIA) was based on the recognition of a pair of antibodies to the protein, − improving the CL sensitivity and selectivity. Since PCSK9 is slightly expressed in serum, a sequence-independent nuclease named T7 exonuclease was employed to further improve the sensitivity. , …”

A methylene blue-functionalized DNA concatemer for the construction of a turn-off fluorescent immunosensor for the sensitive detection of carcinoembryonic antigen

“…7 Among them, fluorescence sensing has become popular and received wide attention due to its simple operation, fast response, low cost, and high sensitivity. Furthermore, many modern signal amplification methods have been developed such as polymerase chain reaction (PCR), 8 enzyme catalysis, 9 catalytic hairpin self-assembly (CHA), 10 and chain hybridization reaction (HCR), 11 and all of them were used to improve the sensitivity of biosensors. However, PCR and enzyme-catalyzed reaction conditions are harsh, and the operation process is complex and costly.…”