Biosensors & bioelectronicsJun 21, 2026

Using modified carbon nanotubes to improve CRISPR/Cas12a detection of hepatitis C virus with a DNA-based signal amplifier

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jun 22, 2026

Abstract

Detection limits of 0.23 fM for fluorescence and 4.1 fM for colorimetric modes were achieved in HCV RNA detection.

A dual-modal biosensing platform integrates fluorescence and colorimetric readouts for HCV RNA detection.
The presence of HCV RNA triggers a catalytic assembly circuit that generates a Y-shaped DNA structure.
This structure activates CRISPR/Cas12a nuclease activity, leading to signal amplification for sensitive detection.
Hemin-binding aptamer-functionalized single-walled carbon nanotubes enhance sensitivity by quenching fluorescence and oxidizing a dye.
In the presence of target HCV RNA, signal interference occurs, allowing for fluorescence recovery and reduced dye oxidation.

Simplified

Hepatitis C virus (HCV) is a major etiological agent of liver diseases and remains a serious global health threat. Herein, we report a dual-modal HCV biosensing platform for ultrasensitive HCV RNA detection by integrating near-infrared fluorescence and colorimetric readouts. In this system, the presence of HCV RNA initiates a catalytic assembly circuit (CAC) that forms a Y-shaped DNA structure, exposing two double-stranded DNA activators with complete protospacer adjacent motif (PAM) to trigger CRISPR/Cas12a nuclease activity. The combination of CRISPR/Cas12a-driven signal amplification and hemin-binding aptamer-functionalized single-walled carbon nanotubes (HeApt-SWCNTs) enables highly sensitive target quantification. Upon exposure to hydrogen peroxide (HO), ferric ion in hemin catalyzes a Fenton-like reaction, generating hydroxyl radicals (·OH) that quench SWCNT fluorescence and oxidize 3,3',5,5'-tetramethylbenzidine (TMB) to its blue oxidized form (oxTMB). In the presence of target HCV RNA, CRISPR/Cas12a-mediated HeApt cleavage inhibits ·OH generation, resulting in SWCNT fluorescence recovery and suppresses TMB oxidation. Under optimal conditions, detection limits of 0.23 fM and 4.1 fM are achieved for the fluorescence and colorimetric modes, respectively. This integrated CAC-Cas12a-HeApt-SWCNTs (CCHS) biosensing strategy offers high specificity, dual-mode signal reliability, and broad potential for early diagnosis of HCV and other RNA viruses. 2 2

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Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

PubMed

Using modified carbon nanotubes to improve CRISPR/Cas12a detection of hepatitis C virus with a DNA-based signal amplifier

Abstract

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