Reverse transcription loop-mediated isothermal amplification kit for swine epidemic encephalitis B virus and application of kit

Abstract

The invention discloses a visualized reverse transcription loop-mediated isothermal amplification kit for a swine epidemic encephalitis B virus and application of the kit. The kit comprises RT-LAMP primers, two reaction buffer solutions, an EM, a fluorescent visual detection reagent, ultrapure water and a swine epidemic encephalitis B virus RNA template; the RT-LAMP primers comprise the outer primers F3 and B3, the inner primers FIP and BIP and the loop primers LF and LB, and the application includes detection of a swine epidemic encephalitis B lesion tissue sample and toxic mosquitoes through the kit. Specificity detection and sensitivity detection prove that the reverse transcription loop-mediated isothermal amplification kit can monitor a reaction in real time and quantitatively detect out the copy number of the swine epidemic encephalitis B virus to quickly and accurately obtain a detection result, and convenience is brought to easy, convenient, quick and reliable detection of the swine epidemic encephalitis B virus.

Description

technical field [0001] The invention relates to the technical field of microbial detection, in particular to a fast, visualized and real-time quantitative detection of porcine epidemic Japanese encephalitis virus loop-mediated isothermal amplification kit and its application. Background technique [0002] Japanese encephalitis ( Japanese encephalitis, JE ) is an insect-borne zoonotic natural foci infectious disease caused by the Japanese encephalitis virus (JEV), which mainly damages the central nervous system. In temperate tropical regions, JEV is mainly transmitted through the mosquito-waterfowl or pig-waterfowl route all year round. At present, there are 3 billion people in the world living in JE endemic areas, which are mainly prevalent in the Far East and Southeast Asian countries and regions, mainly in Asia, and China, India and Southeast Asian regions are more prevalent. The prevalence of JE was first reported in Japan in the 1930s. In the summer of 1924, Japanese ...

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Problems solved by technology

[0005] The purpose of the present invention is to provide a simple, quick and accurate kit for detecting porcine encephalitis Japanese encephalitis virus at the grassroots level in order to solve the problems of difficult, time-consuming and expensive instruments for detecting porcine encephalitis Japanese encephalitis virus at the grassroots level. The technical scheme used for realizing the object of the present invention is: a kind of kit of porcine epidemic Japanese encephalitis virus reverse transcription loop-mediated isothermal amplification, which kit includes RT-LAMP primers, 2× reaction buffer, EM, fluorescent Visual detection reagent, ultrapure water and Japanese encephalitis virus RNA template; the RT-LAMP primers include outer primer F3 (SEQ ID NO: 1) and B3 (SEQ ID NO: 2), inner primer FIP (SEQ ID NO: 3) with BIP (SEQ ID NO: 4) and loop primer LF (SEQ ID NO: 5) with LB (SEQ ID NO: 6):

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