CRISPR-Cas13 a-based bovine viral diarrhea virus detection method

A technology of bovine viral diarrhea and virus, which is applied in the field of detection of bovine viral diarrhea virus, can solve the problems of high cost, difficult to deal with on-site and large-scale detection, and high probability of false positive, achieving high application value and high accuracy , the effect of rapid detection

Active Publication Date: 2020-11-24
SHIHEZI UNIVERSITY
View PDF4 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional BVDV methods almost all require sophisticated instruments and high costs, which are difficult to deal with on-site and large-scale detection, and the probability of false positives is relatively high (Adams M J, Hendrickson R C, Dempsey D M, et al.Tracking the changes in virustaxonomy[J].Archives of virology,2015,160(5):1375-1383.)

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • CRISPR-Cas13 a-based bovine viral diarrhea virus detection method
  • CRISPR-Cas13 a-based bovine viral diarrhea virus detection method
  • CRISPR-Cas13 a-based bovine viral diarrhea virus detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0027] The present invention will be further described in detail below in conjunction with specific embodiments, and the given examples are only for clarifying the present invention, not for limiting the scope of the present invention.

[0028] The inventor expresses and purifies LwCas13a protein with LwCas13a as the research center, and at the same time, designs a specific crRNA according to the 5'UTR conserved region of the reported BVDV gene sequence. The RNA obtained from BVDV was used to verify that Cas13a also has RNA-cleaving activity in vitro, and can specifically and accurately detect BVDV, and the invention was completed on this basis.

[0029] Step 1: Primer design for the detection method of bovine viral diarrhea virus based on CRISPR-Cas13a

[0030] 1. LwCas13a gene primer design

[0031] The full-length gene sequence of pC013-Twinstrep-SUMO-LwCas13a was obtained on addgene, and the full-length primers of LwCas13a protein were designed with Premier5 software. Th...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Sensitivityaaaaaaaaaa
Login to view more

Abstract

The invention discloses a primer for detecting bovine viral diarrhea virus, a kit prepared by using the primer and used for detecting BVDV, and a CRISPR-Cas13 a-based bovine viral diarrhea virus detection method. The primer is 5'-GGGGAUUUAGACUACCCCAAAAACGAAGGGGACUAAAACGCCAUCCAACGAACUCACCACUGUUGCU-3'; the kit at least comprises the primer; a sample to be detected is added into the kit for detection. According to the CRISPR-Cas13 a-based bovine viral diarrhea virus detection method, a CRISPR-Cas13a system is successfully applied to nucleic acid detection of the BVDV for the first time, and whether the BVDV is contained or not can be identified only by adding a micro-scale sample. The CRISPR-Cas13 a-based bovine viral diarrhea virus detection method has high specificity and sensitivity. The method provides a wide prospect for early diagnosis of the BVDV.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a detection method for bovine viral diarrhea virus based on CRISPR-Cas13a. Background technique [0002] Bovine viral diarrhea (mucosal disease) is a contagious disease caused by Bovine Viral Diarrhea Virus (BVDV). It mainly infects cattle, but also other organisms such as pigs, sheep, and camels. wide. BVDV is a positive-sense ribonucleic acid (RNA) virus belonging to the family Flaviviridae and the genus Pestivirus. Infection can lead to a variety of clinical manifestations, including respiratory, gastrointestinal and reproductive (various forms) disorders ( B, Roch F F, Richter V, et al. A meta-analysis of bovine viral diarrhea virus (BVDV) prevalences in the global cattle population [J]. Scientific reports, 2018, 8(1):1-15.). Among them, acute or chronic mucosal diseases, diarrhea, growth and reproduction disorders, immunosuppression and persistent infection...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/682C12N15/11
CPCC12Q1/701C12Q1/682C12Q2563/107C12Q2521/327
Inventor 倪伟胡圣伟姚瑞李村院李晓悦徐越仁
Owner SHIHEZI UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap