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.
Highlights
Bovine Viral Diarrhea Virus (BVDV) is a high-prevalence viral of cattle, affecting herds worldwide and leading to significant economic losses in the cattle industry [1]
The results showed that the fluorescence intensity of BVDV virus and BVDV ribonucleic acid (RNA) detection was significantly higher than that of HEK293T cells and MDBK cells and blank controls
The CRISPR-Cas13a has had a significant impact in probing molecular detection based on genome editing and is promising to provide new strategies in the development detection of RNA viruses
Summary
Bovine Viral Diarrhea Virus (BVDV) is a high-prevalence viral of cattle, affecting herds worldwide and leading to significant economic losses in the cattle industry [1]. The disease results in multiple clinical symptoms in the respiratory system, digestive tract, and reproductive system [3,4,5]. Recessive infections of BVDV result in consequences of abortion, which is a key feature in the epidemiology of this disease [6]. Control options for reducing BVDV infections were based on initial screening for serological evidence of persistently infected animals [7]. There are many methods of detection for BVDV virus, including traditional serological neutralization test, immunohistochemical method, ELISA, and, recently, PCR technique [8]. Traditional methods of BVDV are costly and need precise instruments [9], so it is very urgent to establish a fast, accurate, efficient, and visual method to detect BVDV
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