菌物学报2026，Vol.45Issue(9)：95-105,11.DOI:10.13346/j.mycosystema.260042
基于RPA-CRISPR/Cas12a的小麦矮腥黑粉菌快速检测技术
RPA-CRISPR/Cas12a-based rapid detection for Tilletia controversa
摘要
Abstract
Wheat dwarf bunt disease(DB)is a major quarantinable disease caused by the infection of Tilletia controversa(TCK).Accurate and rapid detection of TCK is crucial for port quarantine and disease prevention and control.In this study,a rapid visual detection system combining recombinase polymerase amplification(RPA)with CRISPR/Cas12a was established by targeting a highly conserved gene fragment on the TCK genome(accession No.LWDE02001036.1).This system could complete the detection of the target pathogen within 20 minutes under a constant temperature of 37℃,with the detection sensitivities of 200 fg/μL and 10¹ copies/μL for TCK genomic DNA and recombinant plasmids,respectively,and the minimum detectable limit of 40 teliospores.The detection results can be directly read by the color reaction of nucleic acid test strips.With the advantages of high specificity,high sensitivity and rapidity,this system provides technical support for TCK quarantine at grassroots ports.关键词
快速检测/重组酶聚合酶等温扩增/CRISPR/Cas12a检测体系/小麦矮腥黑粉菌Key words
rapid detection/recombinase polymerase amplification(RPA)/CRISPR/Cas12a detection system/Tilletia controversa(TCK)引用本文复制引用
何畅,崇钰,黄洁,李媛,高利,王保通,李强..基于RPA-CRISPR/Cas12a的小麦矮腥黑粉菌快速检测技术[J].菌物学报,2026,45(9):95-105,11.基金项目
科技部对发展中国家援助项目(KY202002018) （KY202002018）
陕西省重点研发计划(2025NC-YBXM-049) This work was supported by the Ministry of Science and Technology Aid Project to Developing Countries(KY202002018)and the Key Research and Development Program of Shaanxi Province(2025NC-YBXM-049). （2025NC-YBXM-049）
