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Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Published on: December 23, 2022

CbAgo-enriched Cas12a biosensor for cancer mutations screening.

Xiwen Zhai¹, Biyao Yang¹, Fei Deng²

¹School of Biomedical Engineering, ARC Centre of Excellence in Nanoscale Biophotonics, Faculty of Engineering, UNSW Sydney, NSW, 2052, Australia.

Analytica Chimica Acta

|April 11, 2026

Summary

This summary is machine-generated.

A new system, CbAgo-enriched Cas12a mutation screening system (CECMS), enhances detection of rare DNA mutations in body fluids. This breakthrough improves cancer monitoring and diagnostics by increasing sensitivity for low-frequency variant allele frequencies.

Keywords:

CRISPR-Cas12a CbAgo Circulating tumor DNA (ctDNA)KRAS G12D Variant allele frequencies (VAFs)

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Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Accurate detection of low-frequency DNA mutations in body fluids is critical for cancer monitoring and treatment.
High levels of wild-type DNA in samples can obscure rare mutant signals, hindering sensitive detection.

Purpose of the Study:

To develop a highly sensitive method for detecting low-frequency DNA mutations in biological fluids.
To overcome the challenge of wild-type DNA interference in mutation screening.

Main Methods:

Developed the CbAgo-enriched Cas12a mutation screening system (CECMS).
Integrated CbAgo's single-nucleotide resolution with CRISPR-Cas12a's trans-cleavage activity.
CECMS selectively removes wild-type DNA and enriches target mutant alleles.

Main Results:

CECMS demonstrated up to 100-fold increased sensitivity compared to conventional Cas12a biosensors at 37°C.
Achieved reliable detection of variant allele frequencies (VAFs) as low as 0.01%.
Successfully detected KRAS G12D mutations (a pancreatic cancer marker) at a VAF of 0.1% in spiked serum samples for circulating tumor DNA (ctDNA).

Conclusions:

CbAgo-mediated enrichment significantly enhances the detection of rare single nucleotide variants (SNVs).
CECMS offers high efficiency and ease of use, making it a promising tool for clinical cancer diagnostics.
The system holds strong potential for improved cancer monitoring and patient management.