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

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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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 measures for the treatment of African swine fever and the prevention and control of vaccines. The most important current prevention and control measures for the disease are still quarantine. Once the disease is detected, the animals will be strictly culled and taken harmless processing

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