Related Experiment Video
Updated: Sep 13, 2025
09:03
Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a
Published on: December 23, 2022
2.7K
Ultrasensitive eccDNA Detection for Tumor Diagnostics by Using CRISPR/Cas12a-Coupled Nested PCR.
Gege Liu1, Junlu Wu1, Bin Yang1
1Department of Laboratory Medicine, Shanghai Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China.
Analytical Chemistry
|July 31, 2025
Summary
Extrachromosomal circular DNA (eccDNA) shows promise as a cancer biomarker. A new ultrasensitive method, NPCC, enhances eccDNA detection in liquid biopsies for improved cancer diagnostics.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Extrachromosomal circular DNA (eccDNA) are novel biomarkers for cancer detection.
- Low abundance in biofluids and lack of robust detection techniques hinder clinical application.
Purpose of the Study:
- Screen for tumor-associated eccDNA biomarkers.
- Develop a novel ultrasensitive detection method for eccDNA in liquid biopsies.
Main Methods:
- Developed NPCC (Nested PCR-CRISPR/Cas12a) assay.
- Combined nested PCR for eccDNA amplification with CRISPR/Cas12a for sequence-specific detection.
- Validated with synthetic standards and plasma samples from 88 cancer patients.
Main Results:
- Achieved a limit of detection of 10-6 fM, over 100-fold improvement over conventional PCR.
- Demonstrated no cross-reactivity with linear or genomic DNA.
- Successfully detected tumor-specific eccDNAs, including eccDNA-HCC-1 in hepatocellular carcinoma patients (AUC = 0.8977).
Conclusions:
- NPCC overcomes key technical barriers in liquid biopsy for eccDNA detection.
- Offers a cost-effective, highly sensitive, and specific platform for noninvasive cancer diagnostics.
