# Droplet Digital CRISPR for Nucleic Acid Detection

## TL;DR

Droplet digital CRISPR combines CRISPR's precision with droplet technology to detect nucleic acids with high sensitivity and accuracy.

## Contribution

This review introduces the integration of CRISPR with droplet digital technology for nucleic acid detection and quantification.

## Key findings

- ddCRISPR enables single-molecule resolution and minimizes background interference through picoliter microdroplets.
- Amplification-based and amplification-free detection strategies have been advanced for DNA and RNA biomarker detection.
- Challenges include workflow automation, droplet stability, and assay portability, with future directions involving AI and point-of-care integration.

## Abstract

Droplet digital (dd) clustered regularly interspaced short palindromic repeats (CRISPR) integrates the high sequence specificity of CRISPR‐based nucleic acid detection with the absolute quantification capability of digital droplet microfluidics, offering high sensitivity, precision, and scalability. By partitioning samples into thousands to millions of picoliter microdroplets, ddCRISPR enables single‐molecule resolution and minimizes background interference. This review summarizes the principles of droplet generation, manipulation, and detection in ddCRISPR platforms, as well as recent advances in amplification‐based and amplification‐free detection strategies. Representative applications are highlighted for viral, bacterial, and other DNA/RNA biomarker detection. Current challenges, including workflow automation, droplet stability, multiplexing, and assay portability, are discussed alongside future perspectives such as artificial intelligence (AI)‐assisted analysis, point‐of‐care integration, and high‐throughput multiplexed detection. These insights aim to guide the translation of ddCRISPR technologies from laboratory research to robust, scalable, and accessible diagnostic solutions.

This review outlines recent advances in droplet digital CRISPR technology for nucleic acid detection, combining CRISPR specificity with droplet digital‐based absolute quantification. It summarizes core principles, amplification‐assisted and amplification‐free strategies, and representative DNA and RNA biomarker applications. Current challenges and future opportunities in multiplexing, automation, and portable integration are discussed to support next‐generation molecular diagnostics.