Primers and kit for rapidly detecting African swine fever virus

Abstract

The invention relates to primers and kit for rapidly detecting African swine fever virus. Four primers are provided, the kit comprises the primers for detecting nucleic acid of the African swine fevervirus and 18 [mu]L of a reaction solution, and the reaction solution comprises the following reagents: 1.8 [mu]L of 40 mM KCl, 1.8 [mu]L of 100 mM(NH4)2SO4, 1.8 [mu]L of 80 mM MgSO4, 1.8 [mu]L of 1%Tween-20, 0.9 [mu]L of28 mM dNTPs, 1.8 [mu]L of 8000 U / mL Bst enzyme, 0.18 [mu]L of 60 U specific endonuclease FEN1, 0.9 [mu]L of 1 mM SYBRGREEN fluorochrome, 1.8 [mu]L of 4.0*10<-5> mol / L gold nanoparticles and 0.22 [mu]L of ultrapure water. Compared with the prior art, the primers and kit for rapidly detecting the African swine fever virus have the following advantages: four different specific primers are provided, so that the accuracy of detection results for the African swine fever virus is higher, accurate detection results can be rapidly given in combination with an improved LAMP technology and a microfluidic chip technology, and the purpose of jointing detecting multiple different indexes of the same sample can be achieved.

Description

technical field [0001] The invention belongs to the field of genetic engineering, in particular to primers and a kit for rapidly detecting African swine fever virus. Background technique [0002] African Swine Fever (ASF) is an acute, febrile and highly contagious viral disease in pigs. It is characterized by a short course of disease, but the mortality rate is as high as 100%. The clinical manifestation is fever. , Skin cyanosis, lymph nodes, kidneys, gastrointestinal mucosa bleeding significantly. The World Health Organization (OIE) lists it as a disease that must be notified, and the detection techniques for African swine fever include enzyme-linked immunoassay (ELISA), polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP). , ELISA can only be used to detect whether there are antibodies to African swine fever virus in animals. To be precise, it can only detect pigs that have been infected or vaccinated, but it cannot monitor the African swine ...

AI Technical Summary

Problems solved by technology

The World Health Organization (OIE) lists it as a disease that must be notified, and the detection techniques for African swine fever include enzyme-linked immunoassay (ELISA), polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP). , ELISA can only be used to detect whether there are antibodies to African swine fever virus in animals. To be precise, it can only detect pigs that have been infected or vaccinated, but it cannot monitor the African swine fever epidemic. Very good results; while PCR technology is more classic, but its reaction needs to be cycled in two different temperature zones, which requires high equipment and relatively high cost, and PCR technology only needs 1 pair of amplification primers, which is susceptible to interference and specific Relatively insufficient, and the result time is long, professional operation requirements are high, and there are many steps for micro-dosing; the loop-mediated isothermal amplification method is a new type of nucleic acid amplification method, which is characterized by the design of 6 regions for the target gene 4 kinds of specific primers, under the action of strand-displacing DNA polymerase, amplify at a constant temperature of 60--65°C, and the nucleic acid can be amplified in about 15-60 minutes, and the efficiency can reach 10 9 ~10 10 An order of magnitude, with the characteristics of simple operation, strong specificity, and easy detection of products

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