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Truncated and expressed duck plague virus (DPV) recombinant envelope gI protein and preparation method and application thereof

A technology of duck plague virus and capsule, which is applied in the field of veterinary medicine and biology, can solve the problems of cumbersome operation, unsuitable serum sample detection, limited purity of whole virus antigen, etc., and achieve the effect of high purity and good antigenic reactogenicity

Active Publication Date: 2013-04-24
SICHUAN AGRI UNIV
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  • Summary
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  • Claims
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AI Technical Summary

Problems solved by technology

The classic method is the serum neutralization test, but it is not suitable for the detection of large quantities of serum samples due to its long test cycle, cumbersome operation and poor sensitivity.
EL ISA for DPV antibody detection has the advantages of simplicity, sensitivity, and specificity. Combined with semi-automatic detection equipment, it is suitable for monitoring the antibody level of a large number of ducks and the quarantine needs of grassroots units; however, the ELISA method established in the past uses the whole virus as the coating antigen. , due to the technical limitations of virus culture and purification, the purity of the whole virus antigen is limited, the stability is poor, and it is difficult to standardize, which affects the accuracy of ELISA detection results and limits the large-scale application of the ELISA method (DPV-ELISA) coated with whole virus. scale application

Method used

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  • Truncated and expressed duck plague virus (DPV) recombinant envelope gI protein and preparation method and application thereof
  • Truncated and expressed duck plague virus (DPV) recombinant envelope gI protein and preparation method and application thereof
  • Truncated and expressed duck plague virus (DPV) recombinant envelope gI protein and preparation method and application thereof

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Embodiment 1

[0039] Example 1 Truncated Expression of Duck Plague Virus Recombinant Envelope gI Protein and Its Preparation and Application

[0040] 1. Extraction of DNA

[0041] 1. Proliferation and culture of DPV

[0042] Inoculate the DPV CHv seed virus into the newly grown dense monolayer duck embryo fibroblasts (DEF), discard the virus liquid after adsorption at 37°C for 60 minutes, and then add calf serum with a volume fraction of 2% and 100IU / mL double Antibiotic MEM was used to maintain the nutrient solution, and then cultured at 37°C for 24-48 hours.

[0043] 2. DNA extraction

[0044] 1) Select DEF (100mL cell bottle) with 70% cytopathic effect (CPE) after infection with DPV seed virus;

[0045] 2) Pour off the cell culture medium, add 500 μL of cell lysate, and at the same time add proteinase K (10 mg / mL) to a final concentration of 200 μg / mL, mix gently, and incubate at 37 ° C for 10 minutes;

[0046] 3) Pour the cell suspension into an EP centrifuge tube, wash the lysate r...

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Abstract

The invention belongs to the field of animal medicament and biotechnology and particularly relates to a truncated duck plague virus (DPV) recombinant envelope gI protein. The DNA sequence of the truncated and expressed DPV recombinant envelope gI protein is shown as the SEQ ID No.1, and the truncated and expressed DPV recombinant envelope gI protein has a peptide sequence shown as the SEQ ID No.2. The preparation method of the truncated and expressed DPV recombinant envelope gI protein comprises the following steps: acquiring genes of the truncated and expressed DPV recombinant envelope gI protein, preparing a recombinant prokaryotic expression vector of the truncated and expressed DPV recombinant envelope gI protein, and preparing the truncated and expressed DPV recombinant envelope gI protein. The invention also relates to the application of the truncated and expressed DPV recombinant envelope gI protein to preparation of a DPV antigen or vaccine, in particular to the application of the truncated and expressed DPV recombinant envelope gI protein to the preparation of a DPV ELISA (enzyme-linked immuno sorbent assay) antibody test kit. The truncated and expressed DPV recombinant envelope gI protein has high purity, and the preparation method of the truncated and expressed DPV recombinant envelope gI protein is easy to operate, adopts the truncated and expressed DPV recombinant envelope gI protein to carry out ELISA on a DPV antibody in serum, and is simple, efficient and accurate.

Description

technical field [0001] The invention relates to the fields of animal medicine and biotechnology, in particular to truncated duck plague virus recombinant envelope gI protein and its preparation and application. Background technique [0002] Duck plague (Duck plague, DP), also known as duck viral enteritis (Duck Viral Enteritis, DVE), is caused by duck plague virus (Duck plague virus, DPV). , febrile and septic infectious diseases are one of the main infectious diseases that endanger waterfowl farming in the world. At present, there are two types of vaccines used to prevent duck plague: inactivated vaccines and attenuated vaccines. Among them, the attenuated vaccines have better immune efficacy than inactivated vaccines, and the detection of DPV-specific antibody levels is a reasonable way to evaluate the DPV immune effect and formulate The key to the immunization program. At present, the main methods used to detect DPV antibodies include neutralization test (neutralization...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/115C12N15/45C12N15/70A61K39/155A61P31/14G01N33/569C12R1/19
CPCY02A50/30
Inventor 程安春李丽娟汪铭书陈孝跃
Owner SICHUAN AGRI UNIV
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