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CRISPR-Cas13a-Based Detection for Bovine Viral Diarrhea Virus.

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.

SUBMITTER: Yao R

PROVIDER: S-EPMC8219879 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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CRISPR-Cas13a-Based Detection for Bovine Viral Diarrhea Virus.

Yao Rui R Xu Yueren Y Wang Lang L Wang Dawei D Ren Linchang L Ren Changling C Li Cunyuan C Li Xiaoyue X Ni Wei W He Yanhua Y Hu Ruirui R Guo Tao T Li Yaxin Y Li Lei L Wang Xiaokui X Hu Shengwei S

Frontiers in veterinary science 20210609

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 <i>in vitro</i> was v ...[more]

PMID: 34179152

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