Han Hongjian, Zhang Desheng, Hao Weilin, Liu Anjing, Xia Nengwen, Cui Meng, Luo Jia, Jiang Sen, Zheng Wanglong, Chen Nanhua, Gu Jinguo, Bai Jianfa, Zhu Jianzhong
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China.
Animals (Basel). 2025 Jun 27;15(13):1902. doi: 10.3390/ani15131902.
African swine fever virus (ASFV) causes a highly contagious and lethal hemorrhagic disease and significantly threatens the pig industry. There is no commercially effective vaccine available currently, making the detection of ASFV critical for control and prevention. Previously, we established the CRISPR-LbCas12a and LwCRSIRP-Cas13a visual detections of ASFV, separately, targeting the structural p17 gene D117L. In this study, we performed the parallel detections of ASFV based on the conserved viral protease gene S273R using CRISPR-LbCas12a and CRISPR-LbuCas13a systems. Our results showed that both systems are able to specifically detect ASFV as low as two copies of the S273R gene, and effectively detect clinical samples with minimal DNA purification. The work promotes CRISPR-Cas systems for the application of on-site detection in the field.
非洲猪瘟病毒（ASFV）会引发一种具有高度传染性和致命性的出血性疾病，对养猪业构成重大威胁。目前尚无商业上有效的疫苗，因此ASFV的检测对于控制和预防至关重要。此前，我们分别建立了针对结构p17基因D117L的CRISPR-LbCas12a和LwCRSIRP-Cas13a对ASFV的可视化检测方法。在本研究中，我们使用CRISPR-LbCas12a和CRISPR-LbuCas13a系统基于保守的病毒蛋白酶基因S273R对ASFV进行了平行检测。我们的结果表明，这两种系统都能够特异性地检测低至两个拷贝的S273R基因的ASFV，并在DNA纯化最少的情况下有效检测临床样本。这项工作推动了CRISPR-Cas系统在现场检测领域的应用。
