European and America type pig breeding and syndrome virus antibody identification and diagnosis reagent kit preparation method

Abstract

The invention provides a preparation method of a European American type pig propagation and syndrome virus antibody differential diagnosis Kit. The method constructs the pronucleus expression vector pGEX-6P-1-N aimed at American type PRRSV N protein regarding the gene deficiency of the conserved epitopes 25aa-30aa, 37aa-52aa and 50aa-66aa of European and American type strain in an ORF7 gene of a PRRSV VR-2332 strain. Regarding the amino acid sequence of the European type PRRSV N protein epitopes 2aa-12aa, 40aa-46aa and 67aa-128aa, the method establishes the pronucleus recombination expression vector PQE30-N of the European type PRRSV N protein. The recombination plasmid expresses successfully in the bacillus coli, and an indirect ELISA diagnosis Kit for PRRS antibody detection is prepared through using the expression depurated North American type N protein. The coincidence rates of the antibody Kit and an IDEXX Kit both reach 90 percent, and the depurated European type protein serves as enveloping antigen, thereby optimizing the European type PRRSV antibody detection indirection ELISA method, and laying a foundation for PRRS antibody typing detection.

Description

1. Technical field [0001] The invention relates to veterinary disease diagnosis technology in the field of biotechnology, in particular to a preparation method of a European and American type porcine reproduction and syndrome virus antibody differential diagnosis kit. 2. Background technology [0002] The occurrence and prevalence of PRRS have caused considerable economic losses to the pig industry in my country. But at present, the prevention and control of the disease is still a difficult problem faced by countries all over the world. In order to better prevent and control the occurrence and prevalence of this disease, it is urgent to establish a reliable method suitable for large-scale diagnosis and detection of PRRS. Common diagnostic methods for PRRS infection include cell culture and RT-PCR. However, these diagnostic methods are time-consuming, heavy in workload, and require special instruments and equipment, which are far from meeting the current needs of diagnosing...

AI Technical Summary

Problems solved by technology

The virus structural proteins encoded by ORF7 of both have good antigenicity, and the nucleotide sequence of N protein is conserved within the type, and the difference between the types is large

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