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LAMP kit for detecting hogcholera virus and preparation method thereof

A technology of swine fever virus and kit, which is applied in the field of health inspection, can solve the problems of no LAMP kit for detecting swine fever virus, etc., and achieve the effects of easy large-scale promotion and application, broad market prospect, and simple operation

Inactive Publication Date: 2009-02-04
SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no LAMP kit for the detection of CSFV at home and abroad and its application in the detection of CSFV

Method used

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  • LAMP kit for detecting hogcholera virus and preparation method thereof
  • LAMP kit for detecting hogcholera virus and preparation method thereof
  • LAMP kit for detecting hogcholera virus and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1. Specific DNA sequence search

[0049] The genome sequences of multiple strains of classical swine fever virus were retrieved from the American gene database-GenBank, and the homology analysis was carried out by BLAST software to find specific conserved target sequences for LAMP primer design.

[0050] 2. Design of LAMP primers

[0051] Design primers by special LAMP primer design software

[0052] Primer design V3 (http: / / primerexplorer.jp / elamp3.0.0 / index.html),

[0053] The specific LAMP primers designed are (sorted sequence 1-6):

[0054]

[0055] 3. Establish a detection system

[0056] Set different final concentrations of Mg 2+ (2, 4, 6, 8, 10, mM), dNTP (0, 0.2, 0.4, 0.6, 0.8, 1.0.1.2, 1.4mM), the concentration ratio of internal and external primers (1:1, 1:2, 1:4 , 1:61:8, 1:10, 1:12), at different reaction times (10, 20, 30, 45, 60min) and temperatures (57°C, 60°C, 63°C, 65°C, 68°C) The next reaction, and finally obtain the optimal reaction paramete...

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Abstract

The invention belongs to the field of sanitary examination, and relates to a LAMP kit for testing classical swine fever virus and an establishing method and an application thereof. The kit contains a test system which is composed of the LAMP reaction liquid of six LAMP primers. The tests prove that the kit of the invention has good specificity and sensitivity, fast amplification speed, high efficiency and simple and convenient identification. The test system of the invention can rapidly and conveniently test the classical swine fever virus in high-efficiency, high-specificity and high-sensitivity under the isothermal condition of 65 DEG C without complicate instruments, can better satisfy the spot tests for the classical swine fever virus, provides a novel technical platform for food safety testing, can better meet the urgent requirements for the spot testing of foot and mouth diseases at present, is used for the spot testing of import and export quarantine, food sanitary departments, animal breeding farms, etc, and is easy to be popularized in a wide range.

Description

technical field [0001] The invention belongs to the field of sanitation inspection and relates to a kit for detecting swine fever virus, in particular to a LAMP kit for detecting swine fever virus and its establishment method and application. technical background [0002] Swine fever (Swine Fever, SF), also known as hog cholera virus, is called classical swine fever in Europe. Swine fever was first discovered in the United States in 1833 and has now spread to many countries and regions in the world. According to the data released by the FAO-OIE-World Health Organization (FAO-OIE-WHO) Animal Health Yearbook (1992), there are currently 44 countries and regions with swine fever. Mainly distributed in countries and regions in South America, Europe and Asia. The economic loss caused by swine fever is huge. In the Netherlands, in order to control swine fever in 1983 and 1984, a total of 127 million francs were spent on transporting and destroying infected pigs, disinfecting hou...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 李健熊炜王巧全胡永强邱璐刘俊平
Owner SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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