Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Rapid Visual Detection of African Swine Fever Virus Using CRISPR-LwCas13a Lateral Flow Strip Based on Structural Protein Gene D117L

Version 1 : Received: 13 October 2023 / Approved: 13 October 2023 / Online: 16 October 2023 (08:41:51 CEST)
Version 2 : Received: 5 December 2023 / Approved: 6 December 2023 / Online: 6 December 2023 (05:58:45 CET)

A peer-reviewed article of this Preprint also exists.

Zhang, D.; Jiang, S.; Xia, N.; Zhang, J.; Liu, A.; Deng, D.; Zhang, C.; Sun, Y.; Chen, N.; Kang, X.; et al. Development of Visual Detection of African Swine Fever Virus Using CRISPR/LwCas13a Lateral Flow Strip Based on Structural Protein Gene D117L. Veterinary Microbiology 2024, 293, 110073, doi:10.1016/j.vetmic.2024.110073. Zhang, D.; Jiang, S.; Xia, N.; Zhang, J.; Liu, A.; Deng, D.; Zhang, C.; Sun, Y.; Chen, N.; Kang, X.; et al. Development of Visual Detection of African Swine Fever Virus Using CRISPR/LwCas13a Lateral Flow Strip Based on Structural Protein Gene D117L. Veterinary Microbiology 2024, 293, 110073, doi:10.1016/j.vetmic.2024.110073.

Abstract

African swine fever virus (ASFV) is a large double stranded DNA arbovirus that is highly contagious and seriously endangers the lives of domestic and wild pigs. In the past decade, African swine fever (ASF) has spread in many countries in the Caucasus, Russian Federation, Eastern Europe and Asia, causing significant losses to the pig industry. At present, there is a lack of effective vaccine and treatment for ASF. Therefore, the rapid and accurate detection is crucial for ASF prevention and control. In this study, we have developed a portable lateral flow strip (LFS) detection method mediated by recombinase polymerase amplification (RPA) and DNA enzyme (LwCas13a), which is performed at 37 ℃ and visualized by eyes without the need for complex instruments. This RPA-LwCas13a-LFS is based on the ASFV structural protein p17 gene (D117L), with a detection sensitivity up to 2 gene copies. This method is highly specific and has no cross reactivity to 7 other pig viruses. In the detection of 33 clinical samples, the p17 (D117L) RPA-LwCas13a-LFS had 100% coincidence with conventional quantitative PCR (qPCR). These findings demonstrate the potential of this simple, rapid, sensitive, and specific ASFV detection method for on-site ASFV detection.

Keywords

African swine fever virus (ASFV); LwCas13a; Recombinase polymerase amplification (RPA); Lateral flow strip (LFS); Visual detection

Subject

Biology and Life Sciences, Immunology and Microbiology

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