Duck parvovirus indirect ELISA (enzyme-linked immuno sorbent assay) detection reagent kit

A duck parvovirus and detection kit technology, which is applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of increased feed-to-meat ratio of duck flocks, increased breeding costs, and decreased pass rate of ducks out of the market, achieving strong sensitivity, The effect of high specificity and rapid detection

Inactive Publication Date: 2016-02-03
SHANDONG AGRICULTURAL UNIVERSITY
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease mainly leads to an increase in the feed-to-meat ratio of duck flocks, an increase in breeding costs, and a decrease in the pass rate of ducks for slaughter
Huge economic losses are caused to the meat duck farming industry

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Duck parvovirus indirect ELISA (enzyme-linked immuno sorbent assay) detection reagent kit
  • Duck parvovirus indirect ELISA (enzyme-linked immuno sorbent assay) detection reagent kit
  • Duck parvovirus indirect ELISA (enzyme-linked immuno sorbent assay) detection reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Obtaining of Purified Recombinant Duck Parvovirus VP3 Protein

[0038] A. Extraction of viral DNA: After digesting part of the liver homogenate suspension of affected ducks, the total DNA was extracted by the phenol-chloroform method, and finally the total DNA was extracted with 50 μL ddH 2 O dissolved and stored at -20°C;

[0039] B. Design and synthesis of primers: According to the published complete genome sequence of duck parvovirus, combined with our sequencing results of multiple strains of duck parvovirus isolate VP3, a pair of primers were designed using Primer Premier 5.0 and the restriction enzyme BamHI was added and HindIII restriction site, the nucleotide sequence of F1:5'-CGGATCCAAGGAAGTCACAACGCAGGAT-3' is shown in SEQ ID No.3, and the nucleotide sequence of R1:5'-CAAGCTTGCCATCAGTCTGGTATT-3' is shown in SEQ ID No.4;

[0040] C. PCR amplification: PCR reaction system: 2×EsTaqMasterMix 10 μL, upstream and downstream primers F1 / R1 (20 μmol / L) 1 μL e...

Embodiment 2

[0045] Example 2 Assembly of Duck Parvovirus Indirect ELISA Detection Kit

[0046] Recombinant protein coating: Quantitatively measure the concentration of the purified recombinant protein, take an appropriate amount of recombinant protein and mix it in 10mL 1×carbonate buffer to make the final concentration 1.5ng / L, and then add it to the bottom of the 96-well enzyme plate , 100 μL per well. Cover with sealing film and incubate overnight at 4°C.

[0047] The kit includes the following reagents: 1 piece of 96-well ELISA plate, 20mL of 30×PBST washing solution, 10mL of 1×carbonate buffer (PH=9.6), 2.5g of skim milk powder, 10mL of GoatAnti-DuckIgG-HRP, soluble type One-component TMB substrate chromogenic solution 10mL, stop solution (3mol / LH 2 SO 4 ) 10mL, 4 sheets of sealing film. Store at 2-8°C for 6 months

[0048] The components and proportions of the coating solution, washing solution, blocking solution, sample diluent, chromogenic solution and stop solution are as fo...

Embodiment 3

[0056] The usage method of embodiment 3 duck parvovirus indirect ELISA detection kit

[0057] The specific steps of the method of use are as follows:

[0058] 1. ELISA detection procedure

[0059] (1) Coat the ELISA reaction plate with 150ng / well recombinant duck parvovirus VP3 protein, incubate overnight at 4°C, and wash the ELISA reaction plate 3 times with PBST;

[0060] (2) Seal the ELISA reaction plate with 5g / 100mL skimmed milk powder solution, at 37°C, after blocking for 1 hour, wash the ELISA reaction plate 3 times with PBST;

[0061] (3) Dilute the serum to be tested 1:10 times w / v with 5g / 100mL skimmed milk powder solution, add 100 μL to the ELISA reaction plate, and after 1.5 hours at 37°C, wash the ELISA reaction plate 3 times with PBST;

[0062] (4) Add goat anti-duck IgG-HRP to the ELISA reaction plate at 100 μL / well, and after 1 hour at 37°C, wash the ELISA reaction plate 3 times with PBST;

[0063] (5) Add 100 μL of substrate color development solution to th...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to the technical field of veterinary pathogeny antibody test, in particular to a duck parvovirus indirect ELISA (enzyme-linked immuno sorbent assay) detection reagent kit. The inventor firstly provides a duck parvovirus virus VP3 protein; the amino acid sequence of the protein is shown as SEQ ID No.1; the amino acid sequence expressing the protein is shown as SEQ ID No.2. Study shows that the protein has good immune response original performance, and can be used for DPV (duck plague virus) antibody detection; further, the protein is used for providing the duck parvovirus indirect ELISA detection reagent kit. The reagent kit has the advantages of high speed, high stability, high specificity and high sensitivity, can be used for fast detecting the antibody level of the duck parvovirus virus in the serum, and achieves the obvious promotion effect on the prevention and the control of the disease.

Description

technical field [0001] The invention relates to the technical field of veterinary pathogenic antibody detection and provides an indirect ELISA detection kit for duck parvovirus. Background technique [0002] Duck parvovirus is a kind of parvovirus, which mainly affects ducklings aged 1 to 3 weeks. The main clinical symptoms are diarrhea, dyspnea and soft feet. It occurs all year round and brings great economic benefits to duck breeding. Loss is one of the main diseases in duck breeding, and the morbidity and mortality are also high. [0003] Since March 2015, a disease has appeared in meat ducks in Shandong, Jiangsu, Anhui and other places in my country. It has been verified as duck parvovirus. The disease is characterized by dysplasia of duck beak and protruding tongue. For commercial ducks over the age of one day, the incidence rate of duck flocks is 10%-20%, up to 50%, and the infection rate of large flocks can reach up to 100%, but the disease does not cause the death of...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/015
Inventor 刁有祥陈浩窦砚国冯强郑肖强牛晓宇杨晶于相龙
Owner SHANDONG AGRICULTURAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap