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
作者： Wang, Zhe 1 ; Wang, Yu 4 ; Zhang, Ying 4 ; Qin, Guosong 4 ; Sun, Wenbo 8 ; Wang, Aiping 1 ; Wang, Yanfang 9 ; Zhang, Gaiping 1 ; Zhao, Jianguo 4 ;
作者机构： 1.Zhengzhou Univ, Sch Life Sci, Zhengzhou 450001, Peoples R China
2.Longhu Lab, Zhengzhou 450046, Peoples R China
3.Henan Key Lab Immunobiol, Zhengzhou 450001, Peoples R China
4.Chinese Acad Sci, Inst Zool, Key Lab Organ Regenerat & Reconstruct, Beijing 100101, Peoples R China
5.Univ Chinese Acad Sci, Savaid Med Sch, Beijing 100049, Peoples R China
6.Beijing Inst Stem Cell & Regenerat Med, Beijing 100101, Peoples R China
7.Chinese Acad Sci, Inst Stem Cell & Regenerat, Beijing 100101, Peoples R China
8.Shandong Acad Agr Sci, Inst Anim Sci & Vet Med, Shandong Key Lab Anim Dis Control & Breeding, Jinan 250100, Peoples R China
9.Chinese Acad Agr Sci, Inst Anim Sci, State Key Lab Anim Nutr, Beijing 100193, Peoples R China
10.Peking Univ, Sch Adv Agr Sci, Beijing 100871, Peoples R China
期刊名称：ISCIENCE （ 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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