CRISPR-Cas13a-Based Detection for Bovine Viral Diarrhea Virus

Rui Yao; Yueren Xu; Lang Wang; Dawei Wang; Linchang Ren; Changling Ren; Cunyuan Li; Xiaoyue Li; Wei Ni; Yanhua He; Ruirui Hu; Tao Guo; Yaxin Li; Lei Li; Xiaokui Wang; Shengwei Hu

doi:10.3389/fvets.2021.603919

CRISPR-Cas13a-Based Detection for Bovine Viral Diarrhea Virus

Front Vet Sci. 2021 Jun 9:8:603919. doi: 10.3389/fvets.2021.603919. eCollection 2021.

Authors

Rui Yao¹, Yueren Xu¹, Lang Wang¹, Dawei Wang¹, Linchang Ren¹, Changling Ren¹, Cunyuan Li², Xiaoyue Li¹, Wei Ni¹, Yanhua He^{2

3}, Ruirui Hu¹, Tao Guo¹, Yaxin Li¹, Lei Li¹, Xiaokui Wang¹, Shengwei Hu¹

Affiliations

¹ College of Life Sciences, Shihezi University, Shihezi, China.
² College of Animal Science and Technology, Shihezi University, Shihezi, China.
³ State Key Laboratory of Sheep Genetic Improvement and Healthy Production, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi, China.

Abstract

Bovine Viral Diarrhea Virus (BVDV) is the main pathogen of bovine viral diarrhea disease (BVD), which leads to enormous economic losses in the cattle industry. A sensitive and specific detection for BVDV is advantageous to the control of BVDV. Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have been used for detecting virus RNA. In this study, the expression and purification of LwCas13a protein was optimized and the RNase activity of LwCas13a in vitro was verified. CRISPR-LwCas13a system could detect BVDV virus and BVDV RNA with high specificity and simplicity. The detection limit of the LwCas13a system was 10³ pM, and there were no cross-reactions with HEK293T and MDBK. In summary, a sensitive, specific, and simple nucleic acid detection method based on CRISPR-Cas13a was developed for BVDV. This method provides a new detection strategy for early diagnosis of BVDV.

Keywords: BVDV; CRISPR-Cas13a; LwCas13a; detection; virus.