Primer set, probe kit and detection method for quickly detecting African swine fever virus based on RPA method

Abstract

The invention discloses a primer set, probe kit and detection method for quickly detecting an African swine fever virus based on a RPA method. The primer set and the probe comprise a primer set and aprobe, wherein the primer set comprises a forward primer RPA-F and a reverse primer RPA-R; the total amplification product size of the primer set and the probe is 267bp; and the kit comprises the primer set and the probe. The detection method comprises the following steps of synthesizing the primer set and the probe; and establishing an RPA-LFD detection method. According to the primer set, the probe, the kit and the detection method disclosed by the invention, complex and refined instruments and equipment are not needed, so that the fund investment is reduced; the implement process is simpleand easy to operate, and on-site detection can be performed without specialized detection staff; and the detection is quick and accurate, so that on-site visualized detection of African swine fever can be realized, and the primer set, the probe, the kit and the detection method have important significance to prevention and control of the African swine fever virus.

Description

technical field [0001] The invention belongs to the technical field of African swine fever virus detection, and in particular relates to a primer set, a probe, a kit and a detection method for rapidly detecting African swine fever virus based on an RPA method. Background technique [0002] Recombinase Polymerase Amplification (RPA) is known as a nucleic acid detection technology that can replace PCR. RPA technology mainly relies on three enzymes: recombinases capable of binding single-stranded nucleic acids (oligonucleotide primers), single-stranded DNA binding proteins (SSBs), and strand-displacing DNA polymerases. The protein-DNA complex formed by the combination of recombinase and primer can search for homologous sequences in double-stranded DNA. Once the primers localize a homologous sequence, a strand exchange reaction occurs and DNA synthesis is initiated, exponentially amplifying the target region on the template. The displaced DNA strand binds to the SSB, preventin...

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Problems solved by technology

[0006] Clinical symptoms such as fever, loss of appetite, mental depression, skin cyanosis, dyspnea, vomiting, diarrhea, abortion caused by African swine fever virus and clinical symptoms of classical swine fever, blue ear disease, swine erysipelas, pseudorabies, porcine circovirus, etc Symptoms are similar, and it is difficult to diagnose it based on clinical symptoms alone, and laboratory testing methods must be used, which makes the on-site differential diagnosis of African swine fever difficult

There are many detection methods for African swine fever virus. The mainstream detection methods are PCR and qPCR. These two methods have good specificity, and the qPCR method also has high sensitivity. However, the requirements of the two methods for equipment are relatively high. Strict, requiring sophisticated temperature-changing devices and professional skills, so these two methods can only be tested under laboratory conditions, and take a long time, limiting their application in on-site testing

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