Indirect ELISA detection kit of sheep echinococciosis Eg95 protein antibody

Abstract

The invention discloses an indirect ELISA detection kit of sheep echinococciosis Eg95 protein antibody. The invention is characterized in that the kit comprises an echinococcus granulosus recombinantantigen Eg95 protein precoated ELISA plate, a serum diluent, an enzyme-labeled conjugate diluent, a positive control solution, a negative control solution, an enzyme-labeled conjugate, a washing solution, a substrate developing solution and a stopping solution. During the preparation process of the echinococcus granulosus recombinant antigen, a His tag sequence and a Myc tag sequence are respectively connected to the front and back of the Eg95 gene; then, EcoR I and Xho I are adopted for simultaneous digestion of a PCR product and an eukaryotic expression vector pPIC9K, the target fragment andthe vector are connected after gel extraction, recombinant plasmid pPIC9K-(His-Eg95-Myc) is constructed, the recombinant plasmid is transformed into pichia pastoris GS115 strain, and the recombinantantigen Eg95 is expressed by the induction of methanol and purified by the use of Ni-NTA. The sheep echinococciosis antibody detection kit obtained by the adoption of the dual-tag sequence and eukaryotic expression vector system has strong specificity, high sensitivity and good repeatability, and is easy to operate.

Description

technical field [0001] The invention belongs to the technical fields of genetic engineering technology and diagnostic reagents, and in particular relates to an indirect ELISA detection kit for Echinococcus Eg95 protein antibody. Background technique [0002] Echinococcus is a zoonotic parasitic disease caused by the larvae of the Echinococcus tapeworm (Echinococcus) that parasitizes the liver, lungs and other organs of humans and animals, causing serious harm to human health and the development of animal husbandry. Echinococcosis is distributed worldwide and can not only infect cattle, sheep, pigs, horses, camels and other domestic animals and various wild animals, but also infect humans. It has become an important problem affecting public health and economy worldwide , and thus has received widespread attention. my country is one of the countries with the highest incidence of the disease, and it is the most serious in Xinjiang, Qinghai, Gansu, Ningxia, and northern Xichuan...

AI Technical Summary

Problems solved by technology

[0005] Lightowler et al. developed the Eg95 vaccine against Echinococcus granulosus egg infection. The Eg95 recombinant protein vaccine is currently the only vaccine to control echinococcosis, and clinical trials have confirmed that it can be used for animal immune prevention, but prokaryotic vaccines are mostly used The expression system is used to detect its immune effect, and its specificity and sensitivity are poor. At present, there is no report that the tag sequence is connected before and after the Eg95 gene, and the specificity, specificity, and Therefore, the research and development of an easy-to-operate and highly specific Eg95 vaccine immunological detection kit is of great significance for promoting the popularization and application of Eg95 recombinant protein vaccines

Images