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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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Target Recognition- and HCR Amplification-Induced In Situ Electrochemical Signal Probe Synthesis Strategy for Trace

Aiting Cai1, Luxia Yang1, Xiaoxia Kang1

  • 1School of Public Health, Nantong University, No.9 Seyuan Road, Nantong 226019, China.

Biosensors
|November 10, 2022
PubMed
Summary

A novel electrochemical-DNA (E-DNA) sensor utilizes DNA metallization and hybridization chain reaction (HCR) for sensitive ctDNA detection. This advanced sensor achieves ultra-low detection limits, showing promise for clinical diagnostics.

Keywords:
DNA metallizationE-DNA sensorExo ΙHCRctDNA

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Area of Science:

  • Electrochemistry
  • Biotechnology
  • Molecular Diagnostics

Background:

  • Developing sensitive and selective methods for detecting circulating tumor DNA (ctDNA) is crucial for early cancer diagnosis and monitoring.
  • Existing electrochemical DNA (E-DNA) sensors often face challenges with sensitivity, background noise, and signal amplification.

Purpose of the Study:

  • To construct a highly sensitive E-DNA sensor for trace ctDNA detection.
  • To enhance signal amplification and reduce background noise for improved accuracy.
  • To evaluate the sensor's performance for potential clinical applications.

Main Methods:

  • Fabrication of an E-DNA sensor using DNA metallization for in situ signal reporter generation.
  • Application of hybridization chain reaction (HCR) for signal amplification.
  • Utilizing cyclic voltammetry (CV) to characterize the Ag/AgCl electrochemical process.
  • Incorporation of an enzyme cleavage technique to minimize background signals.

Main Results:

  • The E-DNA sensor demonstrated a wide detection range for ctDNA, from 0.5 fM to 10 pM.
  • Achieved an ultra-low limit of detection (LOD) of 7 aM for ctDNA.
  • Exhibited excellent selectivity, repeatability, stability, and good recovery rates.
  • The sensor showed strong potential for analyzing trace ctDNA in clinical samples.

Conclusions:

  • The developed E-DNA sensor offers superior performance for trace ctDNA analysis.
  • The combination of DNA metallization, HCR, and enzyme cleavage significantly enhances sensing capabilities.
  • The sensor's high sensitivity and reliability support its potential use in clinical diagnostics.