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Related Concept Videos

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Updated: Jan 10, 2026

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A Pre-Amplification-Free Modular Dual-CRISPR System for Enhanced Pathogen Detection Sensitivity.

Tong Su1,2, Tianhui Wei1, Zichun Wang1

  • 1State Key Laboratory of Flexible Electronics (LoFE) & Jiangsu Key Laboratory of Smart Biomaterials and Theranostic Technology, Institute of Advanced Materials (IAM), Nanjing University of Posts and Telecommunications, 9 Wenyuan Road, Nanjing 210023, China.

Analytical Chemistry
|November 25, 2025
PubMed
Summary

A novel dual-CRISPR/Cas12a system coupled with hybridization chain reaction (HCR) enables highly sensitive pathogen nucleic acid detection. This pre-amplification-free approach significantly enhances sensitivity and reduces false positives for diverse pathogen targets.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostics

Background:

  • CRISPR/Cas12a systems offer high specificity for pathogen detection but often require pre-amplification, increasing complexity and risk of errors.
  • Limited sensitivity of single CRISPR/Cas12a systems necessitates nucleic acid amplification, posing challenges like cross-contamination and false positives.

Purpose of the Study:

  • To develop a universal and highly sensitive pathogen nucleic acid detection method without pre-amplification.
  • To enhance the sensitivity and reduce the complexity of CRISPR/Cas12a-based diagnostics.

Main Methods:

  • A modular dual-CRISPR/Cas12a system was designed, incorporating a recognition module and a signal module.
  • The system was coupled with hybridization chain reaction (HCR) and DNA nanostructure self-assembly for signal amplification.
  • The system was tested for detecting various pathogen nucleic acids, including African swine fever virus (ASFV), severe fever with thrombocytopenia syndrome virus (SFTSV), and human papillomavirus type 16 (HPV-16).

Main Results:

  • The dual-CRISPR system achieved attomolar (aM) sensitivity for pathogen nucleic acid detection, surpassing traditional single CRISPR/Cas12a systems by over six orders of magnitude.
  • The system demonstrated high sensitivity and specificity without the need for pre-amplification.
  • The modular system proved versatile, successfully detecting diverse pathogen targets and showing practicality in complex sample environments (e.g., ASFV quality control).

Conclusions:

  • The developed pre-amplification-free dual-CRISPR system offers a significant advancement in sensitive and universal pathogen nucleic acid detection.
  • This approach provides a flexible and robust platform for developing next-generation diagnostic tools with improved performance and reduced complexity.
  • The findings open new avenues for enhancing nucleic acid detection technologies in various applications, including infectious disease surveillance.