Rapid multiplex detection of Echinococcus granulosus and Echinococcus multilocularis using a one-pot RPA-assisted CRISPR-Cas12a/Cas13a assay in a portable multi-tube device.

Authors

Zijian Dong, Yehui Liu, Xiaoxia Wu, Wanli Ban, Li Zhao, Xiaolei Liu, Jing Ding

Published in

Biosensors & bioelectronics. Volume 311. Pages 118857. May 25, 2026. Epub May 25, 2026.

Abstract

Echinococcosis, caused by Echinococcus granulosus and Echinococcus multilocularis, remains a significant zoonotic threat, particularly in pastoral regions where rapid environmental surveillance is essential yet technically constrained. Here, we report a rapid and integrated one-pot recombinase polymerase amplification -assisted, orthogonal CRISPR-Cas12a/Cas13a platform for rapid and specific discrimination of these two species in environmental samples. Coupled with a simplified NaOH-based DNA extraction method, the assay enables a streamlined workflow completed within 60 min, achieving a detection limit of as low as 1 copy/μL without observable cross-reactivity. To facilitate point-of-care deployment, we further developed a low-cost, miniaturized handheld device capable of parallel analysis of up to eight samples with dual-target readout. The platform was validated using field samples, including canine feces, pasture grass, and vegetables, demonstrating complete agreement with quantitative PCR results, with 100% sensitivity and specificity. This integrated CRISPR-based biosensing system provides a robust and field-deployable solution for on-site echinococcosis surveillance and offers a scalable framework for multiplex environmental pathogen detection.

PMID:
42224782
Bibliographic data and abstract were imported from PubMed on 02 Jun 2026.

Read full publication at:
Please sign in to see all details.

Stats

1-terrible, 9-excellent. How would you rate this publication? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.