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In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
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Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
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Hybridization chain reaction-enabled sensing applications: A review.

Shan Huang1, Jianjing Shen1, Xiaoming Ren1

  • 1State Key Laboratory of Materials-Oriented Chemical Engineering, College of Chemistry and Molecular Engineering, Jiangsu Provincial University Key Laboratory of Intelligent Medical Sensing Materials and Devices, Nanjing Tech University, Nanjing, 211816, PR China.

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|December 6, 2025
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Hybridization chain reaction (HCR) offers enzyme-free, isothermal nucleic acid amplification for biosensors. This review explores HCR challenges and future pathways for improved detection technologies.

Keywords:
BiosensorsDNA technologyHybridization chain reactionSignal amplification technique

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Biosensor technologies are rapidly advancing, with signal amplification being crucial for enhanced detection.
  • Hybridization chain reaction (HCR) is a key enzyme-free, isothermal nucleic acid amplification method.
  • HCR utilizes DNA initiators to trigger hairpin probe hybridization, forming extended polymers.

Purpose of the Study:

  • To provide a comprehensive review of Hybridization Chain Reaction (HCR) in biosensing.
  • To systematically examine the technical limitations of current HCR systems.
  • To offer insights into future development pathways for HCR-based biosensors.

Main Methods:

  • Review of existing literature on HCR amplification strategies.
  • Analysis of HCR mechanism, including linear and branched amplification pathways.
  • Evaluation of HCR compatibility with nanomaterials and signal enhancement techniques.

Main Results:

  • HCR demonstrates superior amplification efficiency compared to conventional methods.
  • HCR shows remarkable compatibility with nanomaterials and other signal enhancement strategies.
  • Existing HCR systems face technical limitations that hinder optimal performance.

Conclusions:

  • HCR is a powerful tool for biosensing, offering enzyme-free, isothermal amplification.
  • Addressing current technical limitations is essential for advancing HCR applications.
  • Future research should focus on overcoming these challenges to propel HCR-based biosensor development.