The Nucleic Acid Detection and CRISPR-Based Microfluidic Point-of-Care Biosensing: Research and Applications

Zihao Zhao, Liang Zhao, Xiayan Wang

Prog Chem ›› 2025, Vol. 37 ›› Issue (10) : 1397-1409.

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Prog Chem ›› 2025, Vol. 37 ›› Issue (10) : 1397-1409. DOI: 10.7536/PC20250303
Review

The Nucleic Acid Detection and CRISPR-Based Microfluidic Point-of-Care Biosensing: Research and Applications

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Abstract

Nucleic acid testing is the gold standard and technological cornerstone for the modern diagnosis of pathogenic infections. As a deployable public health surveillance technology, Point-of-Care Testing (POCT) has demonstrated significant value in infectious disease prevention and control, personalized precision medicine, and medical scenarios with limited resources. POCT technology can rapidly provide diagnostic information, significantly improve patient outcomes, and optimize the allocation of medical resources. As an emerging technology, microfluidic chips have become a key component in POCT due to their low reagent consumption, high integration, and automation. By integrating laboratory functions onto a single chip, microfluidic devices have achieved full-process automation of sample processing, signal amplification, and detection, greatly enhancing the efficiency and accuracy of testing. Moreover, when combined with isothermal amplification techniques (such as LAMP) and CRISPR-Cas technology, microfluidic chips can rapidly and sensitively detect pathogens, making them suitable for on-site screening of various infectious diseases. Currently, POCT devices based on microfluidic chips have been successfully applied in the detection of pathogens such as SARS-CoV-2, demonstrating the advantages of speed, portability, and high sensitivity. This review aims to summarize the development of nucleic acid detection and the research progress on the combination of CRISPR-Cas technology and microfluidic chips to explore their current applications and future prospects for POCT.

Contents

1 Introduction

2 Significance of point-of-care nucleic acid testing for pathogens

3 Conventional nucleic acid testing

3.1 PCR-Based nucleic acid testing

3.2 Isothermal-amplification-based pathogen nucleic acid testing

3.3 Other methods

4 CRISPR-Cas biosensor-based nucleic acid testing

4.1 Cas12a-Based nucleic acid detection

4.2 Cas13a-Based nucleic acid detection

4.3 Other CRISPR systems

5 CRISPR-Cas nucleic acid detection on microfluidic chips

5.1 Multiplexed detection on microfluidic chips

5.2 Amplification-free detection on microfluidic chips

5.3 Equipment-free microfluidic POCT for rapid detection

6 Conclusion and prospects

Key words

nucleic acid detection / point-of-care testing (POCT) technology / microfluidic chips / CRISRP-cas biosensing

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Zihao Zhao , Liang Zhao , Xiayan Wang. The Nucleic Acid Detection and CRISPR-Based Microfluidic Point-of-Care Biosensing: Research and Applications[J]. Progress in Chemistry. 2025, 37(10): 1397-1409 https://doi.org/10.7536/PC20250303

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Funding

National Natural Science Foundation of China(22174007)
Outstanding Students Project for undergraduates BJUT, and the Spark Fund for undergraduates, Beijing University of Technology
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