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The hybrid chain reaction is a nucleic acid polymerization reaction without enzyme participation proposed by Dirks and Pierce in 2004.
The target molecule initiates a cascade reaction between several thermodynamically stable DNA fuel chains with specific secondary structures, resulting in ultra-long DNA nanostructures
with cuts.
Signal amplification
of target molecules can be achieved using hybridization chain reactions.
Miao Peng's research group of Suzhou Medical Institute has developed several high-performance sensors for the detection of bioactive molecules based on this technology (Nanoscale, 2022, 14, 612; Part.
Part.
Syst.
Charact.
, 2020, 37, 1900488; ACS Appl.
Mater.
Interfaces, 2019, 11, 41157; Sens.
Actuators, B, 2019, 297, 126788; Chem.
Eng.
J.
, 2018, 353, 305）； Summarizes current advances in the field (ACS Appl.
Mater.
Interfaces, 2021, 13, 38931; TrAC-Trends Anal.
Chem.
, 2017, 94, 1-13）； Several novel hybrid chain reaction assembly methods were proposed (Anal.
Chem.
, 2022, 94, 14755; Anal.
Chem.
, 2020, 92, 12700; Anal.
Chem.
, 2020, 92, 12026, Chem.
Commun.
, 2020, 56, 1175）
。
Recently, the research group has developed a novel linear hybridization system, and combined magnetic nanoparticles and silver nanoparticles for highly sensitive and high-specificity analysis
of target nucleic acids.
Firstly, ferric oxide nanoparticles were prepared for rapid magnetic separation after reaction, while microgold nanoparticles were reduced on their surface for immobilization of sulfhydryl modified DNA probe A
.
Based on the huge specific surface area of the nanomaterial, the fixation
of a large number of probe A chains is realized.
The probe contains four functional regions, two of which are complementary pairs with the target miRNA and can be recognized and acted upon by double-strand-specific nucleases (DSNs) after forming DNA/RNA heterozygous duplexes
。 The DNA in this heterozygous double-stranded is specifically digested and released miRNA for cycling reactions, while yielding two single-stranded probes B and C DNA triangular prism nanostructures
that further act on the electrode interface.
The geometric rigidity of the structure provides suitable space for capturing probes B and C, and the recognition efficiency is significantly improved.
In addition of specific designs of fuel chains H1, H2, H3, H4 Later, the growth of trapezoidal hybrid chain reaction products was realized.
After further capture of silver nanoparticles as electrochemical probes, the miRNA of interest can be quantified
by analyzing the dissolution voltammetry response.
The nanomaterials involved in this work were well characterized (Figure 1), and the corresponding reactions were verified by polyacrylamide gel electrophoresis, AC impedance spectroscopy, cyclic voltammetry, and linear scanning voltammetry (Figure 2).
Under the optimized experimental conditions, the detection limit of 5×10-18 mol/L can be achieved, with good sequence selectivity, and the test has been carried out in the actual clinical samples and achieved good results
.
This work was supported by the Suzhou Basic Research Pilot Project (SJC2021016), and the results were published in Biosensors and Bioelectronics, 2023, 220, 114900 (IF=12.
545).
Link to paper: Figure 1:
of iron oxide nanoparticles and iron oxide@ gold composite nanoparticles.
Fig.
2 Gel electrophoresis images of DNA triangular prism assembly and trapezoidal hybrid chain reaction assembly and electrochemical characterization of each reaction step
