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Single-chain antibody used for detecting canine parvovirus (CPV)

A technology of canine parvovirus and single-chain antibody, which is applied in the field of biomedicine, can solve problems such as high cost or safety, and achieve the effect of simple acquisition method and high detection accuracy

Inactive Publication Date: 2018-10-09
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

IgG and IgY may be effective in the early stages of canine parvovirus disease, but are limited by high cost or poor safety

Method used

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  • Single-chain antibody used for detecting canine parvovirus (CPV)
  • Single-chain antibody used for detecting canine parvovirus (CPV)
  • Single-chain antibody used for detecting canine parvovirus (CPV)

Examples

Experimental program
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Embodiment 1

[0031] Embodiment 1, with reference to Figure 1-9 It is the structural representation of Embodiment 1 of the present invention:

[0032] The technical solutions of the present invention will be described in detail below in conjunction with specific embodiments. :

[0033] (1) Construction of anti-CPV-VLP single-chain antibody library: Mix CPV-VLP and Freund's adjuvant to immunize white Legion chickens by multi-point injection in chest muscles. RNA was reversed to obtain cDNA, and the VH and VL genes were amplified. Assemble scFv by overlap PCR,

[0034] Primers were synthesized according to the sequences in the table, VH and VL genes were amplified, and scFv genes were assembled.

[0035]

[0036] The reaction program of PCR was: pre-denaturation at 95°C for 5 min, denaturation at 95°C for 40 s, annealing at 57°C for 40 s, extension at 72°C for 1 min, 30 cycles, and extension at 72°C for 7 min.

[0037] The scFv was linked to T7 10-3b Vector Arm.

[0038]

[0039] T...

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Abstract

Disclosed is a single-chip antibody used for detecting canine parvovirus (CPV). The single-chip antibody is characterized by establishment of a CPV-VLP (virus like particles) resistant single-chain antibody library, panning of a specific CPV-VLP resistant single-chain antibody and expression of the specific CPV-VLP resistant single-chain antibody. Compared with the prior art, the single-chain antibody has the advantages of high detection accuracy, simple acquisition and the like.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a single-chain antibody for detecting canine parvovirus. Background technique [0002] Canine parvovirus (Canine parvovirus, CPV) is a single-stranded negative DNA virus, containing more than 5200 nucleotides, CPV empty capsid contains VP1, VP2 and VP3 three structural proteins and NS1, NS2 two non-structural proteins (Tsao et al.1991). Among these structural proteins, VP2 accounts for 90% of the viral capsid, and has the function of recognizing host receptors and determining the virus to infect different tissue cells. [0003] Canine myocarditis and hemorrhagic gastroenteritis are different manifestations of CPV infection. Young dogs aged 3-4 weeks are susceptible to acute myocarditis after infection, while adult dogs and dogs older than 10 weeks are susceptible to enteritis syndrome. After CPV infects the cells, the mitochondria of the cells can be swelled, resulting in cell damage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08C07K16/00G01N33/569
CPCC07K16/005C07K16/081C07K2317/622G01N33/56983G01N2333/015
Inventor 张小莺张遥遥李新平杨艳丽原文文
Owner NORTHWEST A & F UNIV
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