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

作者： Min Li*1, Hongkun Qin*1, Yunfei Long1, Meiqin Cheng1, Ling Li1, Aijun Huang2, Nian Wang3, Shuo Duan1 1China-USA Citrus Huanglongbing Joint Laboratory, National Navel Orange Engineering Research Center,Gannan Normal University, 2Department of Biological Sciences,Gannan Normal University, 3Citrus Research and Education Center, Department of Microbiology and Cell Science,University of Florida - Institute of Food and Agricultural Sciences

简介：

Overview

This study addresses the need for rapid and accurate detection of Candidatus Liberibacter asiaticus (CLas), the causative agent of Huanglongbing (HLB) in citrus plants. A new method, referred to as CLas-DETECTR, combines recombinase polymerase amplification with CRISPR-Cas12a technology, allowing for portable and sensitive testing in the field.

Key Study Components

Research Area

• Plant Pathology
• Diagnostics in Agriculture
• Microbial Genetics

Background

• Huanglongbing (HLB) severely impacts citrus production globally.
• CLas early detection is crucial for preventing disease spread.
• Current methods like PCR have limitations, requiring improvement for field applicability.

Methods Used

• Recombinase polymerase amplification combined with CRISPR-Cas12a
• Field samples of citrus leaves were utilized for testing.
• Fluorescent detection technology for result visualization.

Main Results

• CLas-DETECTR showed significantly higher sensitivity than standard PCR.
• Specific fluorescence was observed exclusively in CLas-positive samples.
• The method can yield results within 90 minutes under isothermal conditions.

Conclusions

• The study demonstrates that CLas-DETECTR is a robust and sensitive diagnostic tool for HLB.
• This method can facilitate timely interventions to manage citrus diseases effectively.

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