On-site detection and differentiation of African swine fever virus variants using an orthogonal CRISPR-Cas12b/Cas13a-based assay

文献类型： 外文期刊

第一作者： Wang, Zhe

作者： Wang, Zhe;Wang, Aiping;Zhang, Gaiping;Wang, Zhe;Wang, Aiping;Zhang, Gaiping;Wang, Zhe;Wang, Aiping;Zhang, Gaiping;Wang, Yu;Zhang, Ying;Qin, Guosong;Zhao, Jianguo;Wang, Yu;Zhao, Jianguo;Zhang, Ying;Qin, Guosong;Zhao, Jianguo;Zhang, Ying;Qin, Guosong;Zhao, Jianguo;Sun, Wenbo;Wang, Yanfang;Zhang, Gaiping

作者机构： Zhengzhou Univ, Sch Life Sci, Zhengzhou 450001, Peoples R China;Longhu Lab, Zhengzhou 450046, Peoples R China;Henan Key Lab Immunobiol, Zhengzhou 450001, Peoples R China;Chinese Acad Sci, Inst Zool, Key Lab Organ Regenerat & Reconstruct, Beijing 100101, Peoples R China;Univ Chinese Acad Sci, Savaid Med Sch, Beijing 100049, Peoples R China;Beijing Inst Stem Cell & Regenerat Med, Beijing 100101, Peoples R China;Chinese Acad Sci, Inst Stem Cell & Regenerat, Beijing 100101, Peoples R China;Shandong Acad Agr Sci, Inst Anim Sci & Vet Med, Shandong Key Lab Anim Dis Control & Breeding, Jinan 250100, Peoples R China;Chinese Acad Agr Sci, Inst Anim Sci, State Key Lab Anim Nutr, Beijing 100193, Peoples R China;Peking Univ, Sch Adv Agr Sci, Beijing 100871, Peoples R China

期刊名称：ISCIENCE （ 2023影响因子：4.6； 五年影响因子：5.0 ）

ISSN：

年卷期： 2024 年 27 卷 4 期

页码：

收录情况： SCI

摘要： The African swine fever virus (ASFV) and its variants have induced substantial economic losses in China, prompting a critical need for efficient detection methods. Several PCR-based methods have been developed to discriminate between wild-type ASFV and gene-deleted variants. However, the requirement for sophisticated equipment and skilled operators limits their use in field settings. Here, we developed a CRISPR-Cas12b/Cas13a-based detection assay that can identify ASFV variants with minimal equipment requirements and a short turnaround time. The assay utilizes the distinct DNA/RNA collateral cleavage preferences of Cas12b/Cas13a to detect two amplified targets from multiplex recombinase polymerase amplification (RPA) in a single tube, and the results can be visualized through fluorescent or lateral-flow readouts. When tested with clinical samples in field settings, our assay successfully detected all ASFV-positive samples in less than 60 min. This assay provides a rapid on-site surveillance tool for detecting ASFV and its emerging variants.

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