Double-luciferase-labeled bovine ephemeral fever virus protein compound and application thereof

Abstract

The invention belongs to the technical field of molecular biology, and particularly relates to a dual-luciferase-labeled bovine ephemeral fever virus protein compound and application thereof in detection of bovine ephemeral fever virus infection antibodies and vaccine antibodies. The protein compound disclosed by the invention comprises a firefly luciferase labeled bovine ephemeral fever virus P protein and a renilla luciferase labeled GNS-alpha2-alpha3-beta fusion protein. The protein complex can respectively capture structural protein antibodies generated by bovine ephemeral fever virus infected cattle or vaccine immunized cattle and virus infection specific GNS, alpha2, alpha3 and beta protein antibodies, can realize detection of the two types of antibodies in a single hole by utilizing the difference of two luciferase substrates, and can be used for differential diagnosis of bovine ephemeral fever virus infection and immunization.

Description

technical field [0001] The invention belongs to the technical field of bovine epidemic virus detection, in particular to a double-luciferase-labeled bovine epidemic fever virus protein complex and its application in differential diagnosis of bovine epidemic fever virus infection antibodies and vaccine antibodies. Background technique [0002] Bovine ephemeral fever (BEF) is an acute non-contact infectious disease of cattle, dairy cattle, beef cattle and water buffaloes caused by bovine ephemeral fever virus (BEFV). Because of the epidemic characteristics of the disease, it is also known as three-day fever, temporary fever, rigid disease, etc. The disease can cause reduced milk production and quality of dairy cows, lameness and paralysis of working cattle, and cause significant economic losses to the breeding industry. At present, the disease infected animals include cattle, dairy cows, buffalo, yak, white yak and so on. At present, in the endemic areas of bovine fever, vac...

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Problems solved by technology

[0003] In the early stage, we established an ELISA diagnostic method (ZL 201410244271.1) using G protein as the detection antigen, which can be used to monitor the level of immunity. However, there are G protein antibodies in both vaccinated and naturally infected cattle, so it is impossible to distinguish cattle naturally infected with BEFV or BEFV based on G protein. Antibodies produced by BEF vaccine immunization, in the promotion and application, it was found that suspected sick cattle with a history of vaccination could not be diagnosed by G protein antibody ELISA

At present, it is known that BEFV encodes N, P, M, G and L, α1, α2, α3, β, γ and other proteins, but it is not clear the difference between virus infection and vaccine immunized animals to produce antibodies, which can be used to identify virus antigens of natural infection become a bottleneck in differential diagnosis

Moreover, previous studies have found that the N, P, M, α1, β, γ, and L proteins encoded by BEFV cannot distinguish between BEFV naturally infected cattle and vaccinated cattle. , low or undetectable antibody levels

Both vaccine-immunized cattle and naturally infected cattle produced higher levels of P-protein antibodies. Therefore, using P-protein antibodies as targets cannot distinguish BEFV natural infection of cattle from vaccine immunity.

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