Recombinant protein, coding gene and application thereof and detection kit and detection method for porcine epidemic diarrhea virus antibody

A porcine epidemic diarrhea and recombinant protein technology, which is applied in the field of genetic engineering, can solve the problems of economic loss in pig production and achieve good immunogenicity, intuitive and accurate results, and good specificity

Active Publication Date: 2017-08-04
HEBEI AGRICULTURAL UNIV.
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

In this epidemic, suckling piglets aged within 7 to 10 days were the most seriously affected, with a morbidity rate of up to 100% and a mortality rate of 50% to 90%, which caused huge economic losses to pig production in my country

Method used

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  • Recombinant protein, coding gene and application thereof and detection kit and detection method for porcine epidemic diarrhea virus antibody
  • Recombinant protein, coding gene and application thereof and detection kit and detection method for porcine epidemic diarrhea virus antibody
  • Recombinant protein, coding gene and application thereof and detection kit and detection method for porcine epidemic diarrhea virus antibody

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Experimental program
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Embodiment 1

[0022] 1 Materials and methods

[0023] 1.1 Strains and expression vectors

[0024] E. coli DH5α and BL21 competent cells, prokaryotic expression vector pET-28a (+), are preserved by the Animal Infectious Diseases Laboratory of Hebei Agricultural University.

[0025] 1.2 Main reagents and antibodies

[0026] TRIzol Total RNA Extraction Reagent, dNTPs, Taq DNA Polymerase, and Spin-Column DNA Gel Purification and Recovery Kit were purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.; reverse transcriptase was a product of Promega; restricted nucleic acid endogenous Dicer BamH I and Xho I were purchased from Treasure Bioengineering (Dalian) Co., Ltd.; Ni-Agarose His-tagged protein purification kit was purchased from Beijing Kangwei Reagent Biotechnology Co., Ltd.; protein quantification test kit was purchased from Nanjing Jiancheng Biotechnology Co., Ltd. Institute of Engineering. HRP-goat anti-pig IgG and HRP-goat anti-mouse IgG were purchased from Beijing Suolai...

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Abstract

The invention provides a recombinant protein. The amino acid sequence of the recombinant protein is represented by SEQ ID NO.1. The invention further provides a gene represented by SEQ ID NO.2. The invention further provides application of the protein in detection of a porcine epidemic diarrhea virus antibody. The protein is mainly used as a coating antigen. The invention further provides a kit for detecting porcine epidemic diarrhea virus antibody. The kit contains the recombinant protein, coating liquid, washing liquid, closing liquid, diluent, goat anti-pig IgG marked by horse radish peroxidase and a primer developing solution. The recombinant protein represented by SEQ ID NO.1 is used as the coating antigen for detecting the porcine epidemic diarrhea virus antibody by virtue of an indirect ELISA method and has relatively good specificity, sensitivity and repeatability, and a result is intuitional and accurate.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a recombinant protein, its coding gene, its application, a detection kit and a detection method for porcine epidemic diarrhea virus antibody. Background technique [0002] Porcine epidemic diarrhea (PED) is one of the most important infectious diseases leading to the death of newborn piglets. The disease is caused by porcine epidemic diarrhea virus (PEDV) and is characterized by diarrhoea, vomiting, dehydration and high mortality in suckling piglets. PED first occurred in growing-finish pig herds in the United Kingdom in 1971 and has subsequently occurred in other European countries and Asia. In 1976, PED occurred in Guangdong, Shanghai, Heilongjiang and other places in my country, and then the disease became endemic in domestic pig farms, and PEDV was isolated from sick pigs. In October 2010, newborn piglet diarrhea caused by a mutant PEDV strain broke ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/165C12N15/50G01N33/569
CPCC07K14/005C12N2770/20022G01N33/56983G01N2333/165G01N2469/20
Inventor 宋勤叶姚作俊郭海勇逯纪成孙泰然
Owner HEBEI AGRICULTURAL UNIV.
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