Method of utilizing yolk antibody instead of serum antibody to evaluate infection state of breeder flock

A technology of egg yolk antibody and serum antibody, which is applied in the field of veterinary biology, can solve the problems of only applicable, no detection of egg yolk antibody, and no relationship between serum antibody and egg yolk antibody

Inactive Publication Date: 2012-07-11
SHANDONG AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, all ELISA reagents commercially available to date for the detection of REV, ALV, CAV, and ARV antibodies are only applicable to serum samples.
At present, there is no operating procedure suitable for the detection of egg yolk

Method used

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  • Method of utilizing yolk antibody instead of serum antibody to evaluate infection state of breeder flock
  • Method of utilizing yolk antibody instead of serum antibody to evaluate infection state of breeder flock
  • Method of utilizing yolk antibody instead of serum antibody to evaluate infection state of breeder flock

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Avian reticuloendotheliosis virus antibody (Reticuloendotheliosis Virus Antibody, REV-Ab) detection sample preparation

[0063] Collect fresh eggs; after the eggs are wiped and sterilized by alcohol cotton balls, use burnt tweezers to break open the eggshell (the size of the opening should be inserted with a 1ml tip), and use a 1ml tip to absorb an appropriate amount of yolk; use a kit before testing In the sample diluent, the egg yolk was diluted 200 times, and a two-step dilution method was adopted. Take 10 μl of egg yolk and add it to 90 μl sample diluent, mix well, take out 10 μl from it, add it to 190 μl sample diluent, mix well and set aside.

[0064] Note: Do not dilute the control serum. One tip should be changed for each sample, and the sample should be mixed thoroughly before adding to the antigen-coated well.

[0065] Steps

[0066] Return the reagent to room temperature (18-25°C), shake and mix it before use; take out the antigen-coated plate and mark the...

Embodiment 2

[0081] Detection of Avian Reovirus Antibody (ARV-Ab)

[0082] sample preparation

[0083] 1. Collect fresh eggs; after the eggs are wiped and sterilized by alcohol cotton balls, use burnt tweezers to break open the eggshell (the size of the opening should be inserted with a 1ml tip), and use a 1ml tip to absorb an appropriate amount of yolk; before testing, use The sample diluent in the kit dilutes the egg yolk 400 times, and adopts a two-step dilution method. Take 10 microliters of egg yolk and add it to 190 microliters of sample diluent, mix well, take out 10 microliters from it, add it to 190 microliters of sample diluent, mix well and set aside.

[0084] Note: Do not dilute the control serum. One tip should be changed for each sample, and the sample should be mixed thoroughly before adding to the antigen-coated well.

[0085] Steps

[0086] Return the reagent to room temperature (18-25°C), shake and mix well before use.

[0087] Take out the antigen-coated plate and m...

Embodiment 3

[0100] Avian Leukosis Virus Antibody Test-Subgroup J (ALV-J) Detection Sample Preparation

[0101] Collect fresh eggs; after the eggs are wiped and sterilized by alcohol cotton balls, use burnt tweezers to break open the eggshell (the size of the opening should be inserted with a 1ml tip), and use a 1ml tip to absorb an appropriate amount of yolk; use a kit before testing In the sample diluent, the egg yolk was diluted 250 times, and a two-step dilution method was adopted. Take 10 μl of egg yolk and add it to 115 μl of sample diluent, mix well, take out 10 μl from it, add it to 190 μl of sample diluent, mix well and set aside.

[0102] Note: Do not dilute the control serum. For each sample, a tip should be changed, and the sample should be fully

[0103] well mixed.

[0104] Steps

[0105] Return the reagent to room temperature (18-25°C), shake and mix well before use.

[0106] Take out the antigen-coated plate and mark the position of the tested egg yolk sample on the re...

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Abstract

The invention relates to a method of utilizing yolk antibody instead of serum antibody to evaluate infection state of breeder flock. The method of the invention comprises the following steps: collecting chicken serum and eggs, respectively; respectively detecting antibody levels of serum and yolk by REV, REOV, ALV-J, ALV-AB, CAV and IBDV enzyme linked immunosorbent assay (ELISA) kits from IDEXX company; comparatively analyzing serum and yolk antibody OD value correlation coefficient by SPSS Statistics 17.0 to obtain correlation coefficient of an optimum dilution factor of yolk antibody and a serum antibody OD value according to ELISA detection. The detecting result further indicates that the positive and negative result consistent rate of yolk sample and the serum sample at the optimum yolk dilution factor is 96%-100%. The result of the invention indicates that the serum samples can be replaced by yolk when various infection states of breeder flock are detected by ELISA kits provided by IDEXX for detection with serum samples, so that the infection state of breeder flock can be detected by detecting yolk antibody level.

Description

technical field [0001] The invention relates to a method for evaluating the pathogenic infection status of breeder flocks by yolk antibodies instead of serum antibodies, and belongs to the field of veterinary biology. Background technique [0002] Poultry attenuated vaccines contaminated with exogenous viruses are extremely harmful to the poultry industry. Among them, the most prone to contamination are several viruses (egg-borne viruses) that can be transmitted vertically through chicken embryos, such as avian reticuloendotheliosis virus (REV), avian leukemia virus (ALV), avian infectious anemia virus (CAV) and Reovirus (ARV). At present, the seed viruses used in the production of vaccines have been strictly tested, and once tested, they can be used for many years. This is no longer the main reason why vaccines are contaminated with exogenous viruses. The main source of contamination of the above-mentioned egg-borne viruses in mass-produced attenuated vaccines is the SPF ...

Claims

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Application Information

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IPC IPC(8): G01N33/569
Inventor 崔治中孙淑红赵鹏郑明
Owner SHANDONG AGRICULTURAL UNIVERSITY
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