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Updated: Sep 22, 2025

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

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Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection.

Yangdao Wei¹, Zhenzhen Tao¹, Lu Wan¹

¹State Key Laboratory of Marine Resource Utilization in South China Sea, School of Pharmaceutical Sciences, Hainan University, Haikou, 570228, China.

Biosensors & Bioelectronics

|May 21, 2022

Summary

This summary is machine-generated.

A new Aptamer-based Cas14a1 Biosensor (ACasB) detects live Staphylococcus aureus (S. aureus) without nucleic acid amplification. This highly sensitive and specific biosensor offers rapid on-site detection for food safety and environmental monitoring.

Keywords:

Bacteria diagnostic CRISPR-Cas Staphylococcus aureus

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Published on: April 25, 2025

Area of Science:

Microbiology
Biotechnology
Biosensor Technology

Background:

CRISPR-Cas systems offer versatile pathogen detection via guide RNA modification.
Current CRISPR-based methods often require nucleic acid extraction and amplification, limiting rapid detection.
Sensitive detection of live Staphylococcus aureus (S. aureus) is crucial for food safety and environmental monitoring.

Purpose of the Study:

To develop a novel biosensor for the specific and sensitive detection of live S. aureus.
To create a platform that bypasses the need for nucleic acid extraction and amplification.
To establish an on-site detection method for S. aureus applicable in food safety and environmental contexts.

Main Methods:

Development of an Aptamer-based Cas14a1 Biosensor (ACasB).
Utilized an S. aureus-specific aptamer hybridized with a blocker DNA.
Detection mechanism based on aptamer-S. aureus binding, blocker release, and subsequent Cas14a1 activation leading to a fluorescent signal.
Cas14a1 protein activation by the released blocker binding to sgRNA.

Main Results:

The ACasB demonstrated high specificity and sensitivity, detecting as low as 400 CFU/ml of live S. aureus.
Achieved 100% accuracy in detecting S. aureus in complex samples, comparable to qPCR.
The biosensor functions without requiring nucleic acid extraction or amplification.

Conclusions:

The ACasB provides a sensitive and specific method for detecting live S. aureus.
This aptamer-based Cas14a1 biosensor eliminates the need for nucleic acid amplification, enabling rapid on-site detection.
The ACasB holds significant potential for applications in food safety and environmental monitoring.