Method for fast detecting loop-mediated isothermal amplification of African swine fever viruses

An African swine fever virus, loop-mediated isothermal technology, applied in the field of temperature amplification, can solve the problem of no effective measures, and achieve the effects of low cost, high sensitivity and good specificity

Inactive Publication Date: 2016-04-06
SICHUAN AGRI UNIV
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Problems solved by technology

At present, there is no African swine fever disease in China, and due to the rapid development of world trade, convenient personnel exchanges and other factors, my country is facing huge challenges in the prevention and control of virus invasion
At present, there are no effective mea

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  • Method for fast detecting loop-mediated isothermal amplification of African swine fever viruses
  • Method for fast detecting loop-mediated isothermal amplification of African swine fever viruses
  • Method for fast detecting loop-mediated isothermal amplification of African swine fever viruses

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[0033] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0034] The present invention establishes an African swine fever virus (ASFV) loop-mediated isothermal amplification (loop-mediated isothermal amplification, LAMP) rapid detection method, designs and synthesizes specific LAMP primers for the African swine fever virus K205R gene sequence, optimizes reaction conditions, and analyzes its sensitivity and specificity, the results showed that the established African swine fever LAMP detection method could detect 6 copies / μL of DNA, with high sensitivity, good specificity and stability. This method is simple and fast, and can be completed by reacting in a constant temperature water...

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Abstract

The invention discloses a method for fast detecting loop-mediated isothermal amplification of African swine fever viruses. An LAMP optimal reaction system is prepared from 25 microliters of matter which comprises l microliter of 8U Bst DNA polymerase, 2.5 microliters of a 10*ThermoPol buffer solution, 6 microliters of 2.5 mM each dNTPs, 4 microliters of 25 mM MgCl2, 1 microliter of outer primers F3 and B3 (5 micrometers), 1 microliter of inner primers FIP and BIP (50 micrometers), 1 microliter of a DNA template, 2.5 microliters of 10M bataine, and the balance ddH2O. An LAMP reaction is performed in a constant-temperature water bath kettle, reaction temperature is 63 DEG C, and reaction time is 45 min. The method is extremely high in specificity and sensitivity, fast, efficient, low in cost, easy and convenient to operate, capable of being effectively applied to fast diagnosis of African swine fever viruses, and applicable to field operation of an animal quarantine institution and a livestock farm.

Description

technical field [0001] The invention belongs to the technical field of detection of African swine fever virus, in particular to a method for rapidly detecting ring-mediated isothermal amplification of African swine fever virus. Background technique [0002] African swine fever (ASF) is an acute, highly contagious disease of pigs caused by African swine fever virus (ASFV). Since African swine fever was first discovered in Kenya in 1921, the disease has subsequently broken out on a large scale around the world, causing huge economic losses. The clinical symptoms and pathological changes of African swine fever are similar to those of classical swine fever. It is characterized by high fever, cyanosis of the skin, and obvious bleeding of lymph nodes, kidneys, and gastrointestinal mucosa. The World Health Organization (OIE) classifies it as a class A disease, and my country classifies it as a class I animal disease. At present, there is no African swine fever in China. Due to the...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6844C12Q1/701C12Q2531/119C12Q2563/107
Inventor 王印邬旭龙杨泽晓姚学萍张鹏飞姜睿姣肖璐张博刘亚东冷伊依胡凌林星宇曾相杰罗忠永任梅渗蒙正群
Owner SICHUAN AGRI UNIV
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