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Kit for detecting bovine nodular skin disease virus by eliminating goatpox virus as well as preparation method and application thereof

A sheeppox virus and nodular technology, applied in the field of biology, can solve problems such as poor sensitivity, and achieve the effects of wide application range, accurate and reliable method, and easy operation.

Pending Publication Date: 2020-04-07
BEIJING SENKANG BIOTECHNOLOGY DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Virus isolation and identification takes a long time and needs to be carried out in a certain level of biosafety laboratory. Virus neutralization test is the most specific serological detection method, but because the animal body mainly shows cellular immune response to LSD virus infection, it only produces low levels of Neutralizing antibodies, therefore the method is less sensitive

Method used

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  • Kit for detecting bovine nodular skin disease virus by eliminating goatpox virus as well as preparation method and application thereof
  • Kit for detecting bovine nodular skin disease virus by eliminating goatpox virus as well as preparation method and application thereof
  • Kit for detecting bovine nodular skin disease virus by eliminating goatpox virus as well as preparation method and application thereof

Examples

Experimental program
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preparation example Construction

[0034] A method for preparing a kit for detecting bovine nodular skin disease virus by excluding sheep pox virus, comprising the following steps: aiming at the gene sequence of bovine nodular skin disease virus, selecting a G protein-coupled chemokine receptor gene as a target region , downloaded 37 G protein-coupled chemokine receptor gene sequences of bovine nodular skin disease virus strains, 5 G protein-coupled chemokine receptor gene sequences of sheep poxvirus strains, and 2 goat Poxvirus strain G protein-coupled chemokine receptor gene sequence, a total of 44 sequences, on the basis of multiple sequence alignment, design specific primers and probes that can exclude sheep pox and goat pox virus, said Specific primers include upstream primers and downstream primers, the nucleotide sequence of the upstream primer is shown in SEQ ID NO.1, the nucleotide sequence of the downstream primer is shown in SEQ ID NO.2, the The nucleotide sequence of the probe is shown in SEQ ID NO....

Embodiment 1

[0063] 1. Primer, probe design and construction of bovine nodular skin disease virus pMD20-T-GPCR gene recombinant plasmid

[0064] Primer and probe design

[0065] 1.1 Primer and probe design principles

[0066] In order to ensure the conservation and specificity of the primers and probes, the GPCR gene was selected as the target region, and the GPCR gene sequence of the downstream primer did not match the last two bases of the 3' end of the bovine nodular skin disease virus; When there are multiple bases in the probe that do not match, the upstream primers have the same sequence; when there are two bases that do not match in the probe, the last base at the 3' end of the upstream primer sequence does not match. For details, see figure 1 , to ensure that the primers and probes cannot be amplified simultaneously with the target sequence of the goat pox virus, eliminating the non-specific amplification of the goat pox virus nucleic acid.

[0067] Most of the primer designs in th...

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Abstract

The invention belongs to the technical field of biology, and discloses a kit for detecting bovine nodular skin disease virus by eliminating goatpox virus as well as a preparation method and an application thereof. The kit comprises a specific primer and a probe, wherein the specific primer comprise an upstream primer and a downstream primer, the nucleotide sequence of the upstream primer is as shown in SEQ ID NO.1, the nucleotide sequence of the downstream primer is as shown in SEQ ID NO.2, and the nucleotide sequence of the probe is as shown in SEQ ID NO.3. The kit provided by the invention has the advantages of strong specificity and high sensitivity.

Description

technical field [0001] The invention relates to the technical field of biology, in particular to a kit for detecting bovine nodular skin disease virus by excluding sheeppox virus, a preparation method and application. Background technique [0002] The description of the background technology of the present invention belongs to the relevant technology related to the present invention, and is only used to illustrate and facilitate the understanding of the content of the present invention. prior art on the filing date. [0003] Lumpy skin disease (LSD) is a contagious disease of cattle caused by bovine nodular skin disease virus (LSDV). The incidence of the disease is between 5% and 45%. The case fatality rate can reach up to 10%. The causative agent is bovine nodular skin disease virus, which belongs to the genus Goatpoxvirus of the Poxviridae Vertebratepoxvirinae subfamily, and has a genome homology of up to 96% with sheep poxvirus and goatpoxvirus. The disease is a notifi...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12R1/93
CPCC12Q1/701C12Q1/686C12Q2561/101C12Q2545/113
Inventor 苏莹丁凯杨鹏臧京帅
Owner BEIJING SENKANG BIOTECHNOLOGY DEV CO LTD
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