Detection method for diagnosing Brucella abortus by using recombinant overlapping peptide

A detection method and technology for brucellosis are applied in the field of veterinary drugs and detection for zoonotic diseases, which can solve the problems of difficult detection of brucellosis, and achieve the effects of easy operation, low cost and good quality.

Inactive Publication Date: 2021-08-13
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the deficiencies of the prior art, the present invention provides a detection method for diagnosing Brucella bovine by using recombinant overlapping peptides, which solves the problem that bovine Brucellosis is difficult to detect

Method used

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  • Detection method for diagnosing Brucella abortus by using recombinant overlapping peptide
  • Detection method for diagnosing Brucella abortus by using recombinant overlapping peptide
  • Detection method for diagnosing Brucella abortus by using recombinant overlapping peptide

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Embodiment

[0034] Such as Figure 1-7 As shown, the embodiment of the present invention provides a detection method for diagnosing Brucella bovis using recombinant overlapping peptides, comprising the following steps:

[0035] s1: Screening of antigenic proteins from Brucella outer model protein screening to antigenic protein OMP31;

[0036] s2: Vaccine design, using recombinant overlapping peptide technology to design ROP-OMP31;

[0037] s3: Artificial synthesis, specifically including vector construction, prokaryotic expression and HPLC identification;

[0038] s4: transfection purification, E. coli transformation and protein purification;

[0039] s5: Activity detection of ROP-OMP31, the mice were fed for 3 weeks after intranasal drops, and the activity was detected by acid-fast staining;

[0040] s6: Guinea pig skin test, establish a guinea pig model, and conduct a skin test after 45 days of feeding;

[0041] s7: Cowhide test, establish a bovine brucellosis model, and conduct a s...

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Abstract

The invention provides a detection method for diagnosing Brucella abortus by using a recombinant overlapping peptide, and relates to the technical field of veterinary drugs and detection of zoonosis. The detection method for diagnosing the Brucella abortus by using the recombinant overlapping peptide (ROP-OMP31) comprises the following steps: screening an antigen protein, namely screening the antigen protein OMP31 from Brucella abortus outer mold proteins; vaccine design, namely designing ROP-OMP31 by using a recombinant overlapping peptide technology; artificially synthesizing, namely constructing a vector, performing prokaryotic expression and performing HPLC identification; performing escherichia coli transformation and protein purification; performing activity detection on ROP-OMP31, namely feeding the mouse for 3 weeks after nasal drip, and carrying out staining to detect the activity by using acid-fast staining; performing a guinea pig skin test, establishing a guinea pig model, and performing the skin test after feeding for 45 days; and performing a bovine skin test, establishing a bovine brucellosis model, and performing out the skin test. According to the present invention, the OMP31-ROP design comprises more antigen T cell epitopes; the protein detection purity is higher, and the corresponding quality is better; and meanwhile, compared with a PCR (Polymerase Chain Reaction) technology and an Elisa, the method is easy to operate, safer and lower in cost.

Description

technical field [0001] The invention relates to the field of zoonotic veterinary medicine and detection technology, in particular to a detection method for diagnosing Brucella bovis by using recombinant overlapping peptides. Background technique [0002] Bovine brucellosis is a zoonotic infectious disease that seriously endangers the cattle industry, food safety and public health. In developed countries, bovine brucellosis is well controlled and the incidence rate is low, but it still has a certain prevalence in developing countries; the control strategy of cattle is quarantine and culling, so the detection technology has become an important factor affecting bovine undead bacillosis. important factor controlling the effect. [0003] At present, Elisa and PCR are mainly used as detection methods at home and abroad, but this method is limited by laboratory conditions, and operation in different environments has a great risk of infection. Therefore, the detection of Brucella ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K39/10A61P31/04C07K14/23C12N15/31C12N15/70G01N33/68
CPCC07K14/23C12N15/70G01N33/6866A61K39/098A61P31/04A61K49/0006G01N2333/57
Inventor 王剑
Owner SOUTHWEST UNIV
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