Rapid Visual Detection of African Swine Fever Virus with a CRISPR/Cas12a Lateral Flow Strip Based on Structural Protein Gene D117L

Desheng Zhang; Sen Jiang; Nengwen Xia; Youwen Zhang; Jiajia Zhang; Anjing Liu; Chenyang Zhang; Nanhua Chen; Francois Meurens; Wanglong Zheng; Jianzhong Zhu

doi:10.3390/ani13233712

Rapid Visual Detection of African Swine Fever Virus with a CRISPR/Cas12a Lateral Flow Strip Based on Structural Protein Gene D117L

Animals (Basel). 2023 Nov 30;13(23):3712. doi: 10.3390/ani13233712.

Authors

Desheng Zhang^{1

2

3

4}, Sen Jiang^{1

2

3

4}, Nengwen Xia^{1

2

3

4}, Youwen Zhang^{1

2

3

4}, Jiajia Zhang^{1

2

3

4}, Anjing Liu^{1

2

3

4}, Chenyang Zhang^{1

2

3

4}, Nanhua Chen^{1

2

3

4}, Francois Meurens^{5

6}, Wanglong Zheng^{1

2

3

4}, Jianzhong Zhu^{1

2

3

4}

Affiliations

¹ College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
² Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou 225009, China.
³ Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China.
⁴ Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
⁵ Swine and Poultry Infectious Diseases Research Center, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC J2S 2M2, Canada.
⁶ Department of Veterinary Microbiology and Immunology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK S7N 5E2, Canada.

Abstract

African swine fever virus (ASFV) is a large double-stranded DNA virus that is highly infectious and seriously affects domestic pigs and wild boars. African swine fever (ASF) has caused huge economic losses to endemic countries and regions. At present, there is still a lack of effective vaccines and therapeutics. Therefore, rapid and accurate detection is essential for the prevention and control of ASF. The portable DNA endonuclease (Cas12a)-mediated lateral flow strip detection method (Cas12a-LFS) combined with recombinant polymerase amplification (RPA) has been gradually recognized as effective for virus detection including ASFV. In this study, based on the ASFV structural protein p17 gene (D117L), an RPA-Cas12a-LFS detection method was established. The detection method exhibits a sensitivity of up to two gene copies and has no cross-reaction with nine other swine viruses. Thus, the method is highly sensitive and specific. In 68 clinical samples, the coincidence rate of the p17 strip was 100%, compared to the traditional quantitative PCR (qPCR). In conclusion, we have developed a simple, rapid, sensitive, and specific ASFV visual detection method and demonstrated the potential of on-site detection of ASFV.

Keywords: African swine fever virus (ASFV); Cas12a; D117L; detection; lateral flow strip (LFS); sensitivity and specificity.

Grants and funding

This work was partly supported by the National Key Research and Development Program of China (2021YFD1801201-2), the Jiangsu provincial key R & D plan (BE2020398), the Jiangsu agricultural science and technology independent innovation fund project [CX(21)2035], the National Natural Science Foundation of China (32172867), the 111 Project D18007, and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).