Liquid chip and method for simultaneously detecting antibodies of five poultry vertical transmission diseases

A liquid phase chip and vertical transmission technology, applied in the field of immunochemistry, can solve the problems of weak batch detection ability, long detection time and high cost, and achieve the effects of shortening the detection cycle, high sensitivity and improving efficiency

Active Publication Date: 2017-01-04
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current monitoring of animal diseases still relies on pathogen isolation and culture, serum plate agglutination test, serum neutralization test, indirect immunofluorescence test, enzyme-linked immuno...

Method used

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  • Liquid chip and method for simultaneously detecting antibodies of five poultry vertical transmission diseases
  • Liquid chip and method for simultaneously detecting antibodies of five poultry vertical transmission diseases
  • Liquid chip and method for simultaneously detecting antibodies of five poultry vertical transmission diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: A preparation method of coupled magnetic beads in a liquid chip for simultaneous detection of five poultry vertically transmitted diseases

[0059] In the screening process of a large number of studies in the early stage of the present invention, it was found that only the antigenic protein avian influenza virus NP protein, avian Newcastle disease virus NP protein, avian reovirus δC protein, avian leukosis virus P27 protein, avian reticuloendotheliosis virus The env protein is used in the preparation of the liquid phase chip at the same time, so as to realize the accurate infection of five viruses, namely, avian influenza virus, avian Newcastle disease virus, avian reovirus, avian leukosis virus, and avian reticuloendotheliosis virus. , High sensitivity, rapid, high throughput detection.

[0060] The liquid phase chip is prepared as the above five antigen proteins (avian influenza virus NP protein, avian Newcastle disease virus NP protein, avian reovirus δC p...

Embodiment 2

[0068] Example 2: A preparation method of coupled magnetic beads in a liquid chip for simultaneous detection of five poultry vertically transmitted diseases

[0069] 1) Pretreatment of magnetic beads: Take 5 kinds of blank magnetic beads with different codes, centrifuge them at 10000g for 3min, discard the supernatant, resuspend the blank magnetic beads with activation buffer, vortex for 30s, ultrasonically disperse for 60s, and centrifuge at 12000g 2min and discard the supernatant, repeat 3 times; the activation buffer is 0.1M phosphate buffer saline (PBS), its pH is 6;

[0070] 2) Activation of magnetic beads: Resuspend the magnetic beads after the five pretreatments in the previous step in PBS, vortex for 60 s, and ultrasonically disperse for 60 s, then add 10 μl and 50 mg / mL of nitrogen hydroxysuccinimide sulfonate in sequence Sulfo-NHS and 10 μl, 50 mg / mL carbodiimide (EDC), mixed well, and kept in the dark for 20 minutes at 37°C to obtain activated magnetic beads;

[00...

Embodiment 3

[0076] Example 3: A liquid chip for simultaneous detection of five poultry vertically transmitted diseases

[0077] The liquid chip described in this example contains the coupled magnetic beads prepared in Example 1 or 2, a detection antibody and a reporter molecule.

[0078] The detection antibody is a biotin-labeled anti-chicken IgG antibody.

[0079] The reporter molecule is fluorescent protein or fluorescein-labeled streptavidin.

[0080] The fluorescent protein or fluorescein is selected from one of phycoerythrin, Cy3 and Cy5.

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Abstract

The invention discloses a liquid chip and a method for simultaneously detecting antibodies of five poultry vertical transmission diseases. The liquid chip contains five kinds of fluorescent coding microspheres obtained by coupling magnetic beads of different codes with surface antigens of five poultry vertical transmission diseases, a biotin-labeled detection antibody, streptavidin phycoerythrin and the like. The invention further discloses a preparation method of the liquid chip immune magnetic bead. The liquid chip and the method can simultaneously detect the five poultry vertical transmission diseases with high specificity and high flux, and the whole process is simple in operation, quick and accurate and can realize high-flux detection.

Description

technical field [0001] The invention belongs to the field of immunochemistry, and in particular relates to a liquid phase chip and a method for simultaneously detecting antibodies to five kinds of poultry vertically transmitted diseases. Background technique [0002] The monitoring and purification of important animal diseases in livestock and poultry is an important basis for ensuring the safety of animal-derived food and public health safety. The national standard of "SPF Chicken Microbiology Monitoring" stipulates that there are 19 types of pathogenic microorganisms that need to be monitored in SPF chickens and SPF chicken embryos (including 14 kinds of viral diseases); Clearly put forward the implementation of disease monitoring and purification plans for breeding livestock and poultry farms, giving priority to the prevention and control of highly pathogenic avian influenza, avian Newcastle disease, avian leukemia and other vertically transmitted diseases. However, the ...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/68G01N33/543
CPCG01N33/54326G01N33/56983G01N33/6854G01N2333/08G01N2333/11G01N2333/125G01N2333/15G01N2469/20
Inventor 张建峰孙俊颖李林林刘志成
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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