一种基于RT-RPA-CRISPR / Cas13a的多主棒孢霉菌可视化快速检测方法

By combining RT-RPA-CRISPR/Cas13a technology with fluorescence and colloidal gold test strips, the problem of rapid field detection of *Cercospora pumilus* in durian has been solved, achieving highly sensitive and low-cost visual detection.

CN122405892APending Publication Date: 2026-07-17CHINESE ACAD OF INSPECTION & QUARANTINE +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
CHINESE ACAD OF INSPECTION & QUARANTINE
Filing Date
2026-06-15
Publication Date
2026-07-17

AI Technical Summary

Technical Problem

Existing technologies cannot meet the requirements for rapid, simple, and low-cost field detection of *Cercospora edulis* in durian. Traditional methods rely on specialized equipment and personnel, which cannot meet the needs of on-site testing.

Method used

Using RT-RPA-CRISPR/Cas13a technology, combined with fluorescence detection and colloidal gold test strip detection, a rapid and visualized detection of *Cercospora multipathata* was achieved through specific primer pairs and crRNA.

Benefits of technology

It enables rapid and visual detection of *Cercospora rubrum* in durian, with high sensitivity, strong specificity, and simple operation, making it suitable for field testing and reducing dependence on equipment and personnel.

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Abstract

本发明公开了一种基于RT‑RPA‑CRISPR / Cas13a的多主棒孢霉菌可视化快速检测方法，属于植物病原菌检测技术领域。该检测方法包括：（1）以待测样品基因组DNA为模板，采用SEQ ID NO.4‑5所示的引物对进行逆转录重组酶聚合酶扩增，获得扩增产物；（2）将扩增产物加入CRISPR / Cas13a反应体系中进行切割反应；该体系包含Cas13a蛋白、SEQ ID NO.2所示的crRNA以及报告分子；（3）通过检测报告分子产生的信号判断待测样品中是否存在多主棒孢霉菌。该方法特异性强、灵敏度高，且通过便携式试纸条直接观察，具有操作简便、快速和节约成本的特点，为田间检测提供一种高效的方法。
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