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Biology
Detección de Candidatus Liberibacter asiaticus desplegable en campo mediante amplificaci...
Detección de Candidatus Liberibacter asiaticus desplegable en campo mediante amplificaci...
JoVE Journal
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JoVE Journal Biology
Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a

Detección de Candidatus Liberibacter asiaticus desplegable en campo mediante amplificación de polimerasa recombinasa combinada con CRISPR-Cas12a

Full Text
3,230 Views
09:03 min
December 23, 2022

DOI: 10.3791/64070-v

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

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Please note that some of the translations on this page are AI generated. Click here for the English version.

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.

Frequently Asked Questions

What is Huanglongbing (HLB)?
HLB is a destructive disease affecting citrus plants, caused by Candidatus Liberibacter asiaticus.
How does CLas-DETECTR work?
It combines recombinase polymerase amplification with CRISPR-Cas12a for sensitive and rapid detection of CLas.
What advantages does the CLas-DETECTR method offer?
It provides a portable, fast, and highly sensitive approach for detecting CLas in field conditions.
How long does the detection process take?
The complete testing can be performed in about 90 minutes.
What was the specificity of the method?
Only CLas-positive samples showed green fluorescence, indicating high specificity.
Can this method be used in different environmental conditions?
Yes, the isothermal condition makes it applicable in various field situations.

En este trabajo, se desarrolló un método de detección rápido, sensible y portátil para Candidatus Liberibacter asiaticus basado en la amplificación de la polimerasa recombinasa combinada con CRISPR-Cas12a.

Huang long bing, abreviado como HLB. Es una enfermedad devastadora de los cítricos en todo el mundo. En los países más productores de cítricos, como China y los Estados Unidos, el HLB es causado por la bacteria restringida en floema Candidatus Liberibacter asiaticus, abreviada como CLas.

La detección temprana de CLas fue importante para los productores, facilitando la intervención temprana y previniendo la propagación de la enfermedad. Así que aquí desarrollamos un nuevo método para el diagnóstico rápido y portátil de HLB, combinado con la amplificación de polimerasa recombinasa y el sistema CRISPR-Cas12a. Lo llamamos CLas-DETECTR.

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