RPA primer, reagent and kit for fast detecting BLV (bovine leukemia virus)

A bovine leukemia and provirus technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of restricted use and promotion, and achieve the effect of fast detection speed and low equipment requirements

Inactive Publication Date: 2018-07-27
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] However, PCR and nested PCR methods also have disadvantages, such as the need for specialized techniques and equipment, and are only suitable for laboratories with good equipment, which limits their use and promotion in grassroots laboratories

Method used

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  • RPA primer, reagent and kit for fast detecting BLV (bovine leukemia virus)
  • RPA primer, reagent and kit for fast detecting BLV (bovine leukemia virus)
  • RPA primer, reagent and kit for fast detecting BLV (bovine leukemia virus)

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0036] The present invention utilizes the RPA detection primer of visualization, fast detection BLV proviral nucleic acid to obtain mainly through the following process:

[0037] 1. Extraction and separation of BLV-infected bovine peripheral blood mononuclear cells (PBMC) DNA

[0038] 2. The synthesis of BLV env gene and the construction of its recombinant plasmid and the extraction of nucleic acid

[0039] 3. Determination of the copy number of recombinant plasmid of BLV env gene

[0040] 4. Design and synthesis of BLV RPA primers

[0041] 5. Preparation and operation steps of BLV RPA reaction system

[0042] 6. Establishment and operation steps of BLV RPA-LFA detection method

[0043] 7. Judgment of BLV RPA-LFA detection method results

[0044] 8. Specificity evaluation of BLV RPA detection primers

[0045] 9. Sensitivity evaluation of BLV RPA detection primers

[0046] 1. Extraction and separation of BLV-infected bovine peripheral blood mononuclear cells (PBMC) DNA

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Abstract

The invention discloses an RPA primer, reagent and kit for fast detecting BLV (bovine leukemia virus). The RPA primer is characterized in that the DNA sequence of an upstream primer is <210>2; the DNAsequence of a downstream primer is <210>3. An RPA-LFA technology is used for building a BLV infection visual fast detection system; through the detection of a BLV nucleic acid sample, the result proves that the screened BLV RPA visual detection primer has high specificity and sensitivity. Compared with the prior art, the invention provides a fast, simple, convenient and accurate detection reagentfor the bovine BLV infection; particularly, the advantages of low requirement on the instrument equipment, need of only one constant-temperature water bath pot, fast detection speed (about 40 min), visual detection result, direct observation by naked eyes and the like are realized.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to an RPA primer for visually detecting bovine leukemia provirus nucleic acid and an application thereof, in particular to an RPA primer, a reagent and a kit for rapidly detecting bovine leukemia provirus. technical background [0002] Bovine leukemia (BL), also known as endemic bovine leukemia, is a chronic neoplastic disease of cattle caused by bovine leukemia virus (BLV), characterized by malignant proliferation of lymphoid cells and progressive cachexia. and higher mortality rates. The disease was first discovered in Germany in 1878, and it was not until 1969 that Miller and others in the United States isolated the virus from sick cattle. Subsequently, the disease was discovered in Denmark, Japan, France, South Korea, Russia, Italy, Chile, Poland, Iran, Thailand, Australia, Belgium, Uruguay and other countries and regions, and the infection rate in some areas ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/702C12Q1/6844C12Q2521/507C12Q2522/101
Inventor 乔军孟庆玲张星星张国武李杰王力霞蔡扩军乔梦凡伍晔晖李静刘昱成陈双庆王熙凤
Owner SHIHEZI UNIVERSITY
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