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Indirect ELISA method for detecting duck circovirus antibody

A duck circovirus, antibody technology, applied in the direction of viral peptides, botany equipment and methods, biochemical equipment and methods, etc.

Inactive Publication Date: 2010-08-18
SHANDONG AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Since there is no feasible method for in vitro proliferation and culture of DuCV, the isolation and identification of the virus cannot be completed in the laboratory.

Method used

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  • Indirect ELISA method for detecting duck circovirus antibody
  • Indirect ELISA method for detecting duck circovirus antibody
  • Indirect ELISA method for detecting duck circovirus antibody

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Embodiment Construction

[0021] The gist of the invention is: use pET-32a Prokaryotic Expression Systems to construct a prokaryotic expression vector of part of the Cap gene, purify the expressed recombinant protein, establish an ELISA method, and preliminarily apply it in clinic.

[0022] 1DuCV Cap gene amplification and construction of expression vector

[0023] A pair of primers (5′-ATAGAATTCTTTTTCAGTAGTGACGTT-3′; 5′-TACCTCGAGACTAGAACCCGGTGAA-3′) were designed according to the sequence of the Cap gene registered in GenBank. Using the preserved plasmid DNA as a template, PCR amplifies part of the target gene in Cap, with a size of about 684bp. The PCR product was connected to the pMD18-T vector, identified as a positive clone by enzyme digestion, and named pMD-Cap. The recombinant plasmid pMD-Cap was digested with EcoR I / Xho I to obtain the Cap gene, cloned into the pET-32a(+) vector, identified by EcoR I / Xho I digestion, the positive clone was named pET-32a-Cap, and sent to Sequenced by Shanghai ...

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Abstract

The invention relates to an indirect ELISA method for detecting a duck circovirus (DuCV) antibody. The method uses a pET-32a(+) protokaryon to express the carrier, constructs a gene engineering bacterium BLpET-32a-Cap which can express the partial capsid protein (Cap) of the DuCV, and coats an ELISA plate with the expressed recombinant protein which is subject to purification and renaturation to detect the antibody level of DuCV in the duck blood serum. The result shows that the method has the characteristics of good repeatability and high specificity, and can be used for serological research on the DuCV.

Description

(1) Technical field [0001] The invention relates to an indirect ELISA method for detecting duck circovirus antibody, which belongs to the production field of veterinary biological products. (2) Background technology [0002] Duck circovirus (Duck circovirus, DuCV) is one of the members of the Circovirus genus (Circovirus) of the Circoviridae family (Circoviridae), and is a class of single-stranded circular DNA viruses without an envelope. Since Hattermann first reported it in Germany, it has been discovered or reported in Hungary, the United States, Taiwan, and mainland China. The clinical symptoms of ducks infected with duck circovirus are mainly poor development, and obvious feather dystrophy, especially in the spine (feathers) of the back, bleeding of many feather rachis, and histopathological examination of the back skin. Heterophilic inflammatory exudation of vesicles and tissues surrounding the vesicles. The general pathology of internal organs shows symptoms of co-in...

Claims

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Application Information

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IPC IPC(8): G01N33/569C12N15/34C12N15/70C07K14/01
Inventor 姜世金刘少宁周恩民
Owner SHANDONG AGRICULTURAL UNIVERSITY
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