A method for visual detection of grapevine leafroll-associated virus based on RT-LAMP-CRISPR / Cas12a

By combining RT-LAMP primers with the CRISPR/Cas12a system and alkaline resin buffer to prepare crude nucleic acid extracts, the problems of simplicity, speed, accuracy, and sensitivity in the detection of grape and broad bean wilt virus were solved, achieving efficient virus detection.

CN121183040BActive Publication Date: 2026-05-26FRUIT TREE INST OF CHINESE ACAD OF AGRI SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
FRUIT TREE INST OF CHINESE ACAD OF AGRI SCI
Filing Date
2025-11-03
Publication Date
2026-05-26

AI Technical Summary

Technical Problem

Existing technologies are insufficient for simple, rapid, accurate, sensitive, and specific detection of grape and broad bean wilt virus. Furthermore, the nucleic acid extraction process from grape leaves is complex and subject to interference from strong acidity and polyphenols and polysaccharides.

Method used

By combining RT-LAMP primers with the CRISPR/Cas12a system and alkaline resin buffer, crude nucleic acid extracts were prepared. Rapid and accurate virus detection was achieved through RT-LAMP amplification and CRISPR/Cas12a detection reactions.

Benefits of technology

It achieves highly sensitive and specific detection of grape and broad bean wilt virus, can distinguish it from other viruses, simplifies the nucleic acid extraction process, and improves detection efficiency.

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Abstract

This invention discloses a visual detection method for grape and broad bean wilt virus (GFabV) based on RT-LAMP-CRISPR / Cas12a. Based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) technology and the CRISPR / Cas12a system, this invention develops a highly efficient visual detection method for GFabV. Furthermore, this invention innovatively uses alkaline resin to prepare crude extracts of grape leaves, providing a novel solution to eliminate interference from excessive chlorophyll and other impurities. Experiments demonstrate that the GFabV visual detection method established by this invention based on RT-LAMP-CRISPR / Cas12a is simple, rapid, accurate, highly sensitive, highly specific, and visually appealing, providing a more convenient and efficient technical means for field diagnosis and forecasting of GFabV in my country.
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