Preparation of duck hepatitis I type virus indirect hemagglutination diagnostic antigen and kit

A virus antigen, duck hepatitis technology, applied in the direction of virus peptides, measuring devices, instruments, etc., can solve the problems of cumbersome, time-consuming, high cost, etc., and achieve the effect of convenient and simple use, simple steps, and fast response speed

Inactive Publication Date: 2009-05-06
陶海静
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is cumbersome, time-consuming, and costly, and cannot be used for rapid detection, and is not suitable for popularization and application at the grassroots level

Method used

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  • Preparation of duck hepatitis I type virus indirect hemagglutination diagnostic antigen and kit
  • Preparation of duck hepatitis I type virus indirect hemagglutination diagnostic antigen and kit
  • Preparation of duck hepatitis I type virus indirect hemagglutination diagnostic antigen and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Preparation of sheep red blood cells

[0035] Take 1 part of healthy young ram's blood plus 1 part of Alfred's solution, mix well, and store in a refrigerator at 4°C for 5 days, so that immature or fragile red blood cells can rupture by themselves. Filter the blood with cotton wool to remove fibrin from the plasma. Take 1 part of the filtered blood, add 9 parts of normal saline, mix well and centrifuge at 1000r / min for 10 minutes, discard the supernatant, remove the white blood cells on the surface, add normal saline to resuspend the blood cell sludge, and wash repeatedly 5 times in this way to remove the white blood cells on the surface of the red blood cells. Adhered impurities were then prepared into 8% erythrocyte suspension with PBS solution containing 0.85% NaCl by mass percentage and pH value of 7.2 for later use.

[0036] The formula of Alsever's Solution is: glucose 2.05g, sodium citrate 0.8g, citric acid 0.055g, sodium chloride 0.42g, add distilled water t...

Embodiment 2

[0057] Example 2 Indirect haemagglutination diagnosis antigen detection of duck hepatitis type I virus Specific detection of duck hepatitis type I virus antibody

[0058] Experiment 1: Dilute duck hepatitis type I virus (DHV-I) antigen stock solution by 1:8 and mix it with equal amount of DHV-I positive serum diluted by 1:64, keep it warm in 37°C water bath for 30min, take the above mixed liquid droplet Put it into the wells of a 96-well hemagglutination plate, 25 μl per well, add dropwise to 6 wells, then add the prepared DHV-I indirect hemagglutination diagnostic antigen, 25 μl per well, mix and let it stand for 30 minutes to observe the agglutination phenomenon; the experimental results are shown in Table 1.

[0059] Experiment 2: Incubate DHV-I positive serum at a ratio of 1:64 in a 37°C water bath for 30 minutes, take the above mixture and drop it into the wells of a 96-well hemagglutination plate, 25 μl per well, a total of 6 wells, and then add the prepared DHV-I I ind...

Embodiment 3

[0070] Example 3 Sensitivity Detection of Duck Hepatitis Type I Virus Indirect Hemagglutination Diagnosis Antigen

[0071] Get 10 parts of chicken serum immunized with DHV-I antigen and detect the DHV-1 antibody level with the prepared DHV-1 diagnostic antigen according to the method of the indirect hemagglutination test introduced earlier; meanwhile, these 10 parts of serum are detected by the method of agar diffusion test The level of DHV-I antibody in order to compare its sensitivity. The test results are shown in Table 3;

[0072] table 3

[0073]

[0074]

[0075] Test results: It can be seen from the table that the sensitivity of the indirect hemagglutination test (IHA) is much higher than that of the agar diffusion test (AGP), which is 32 to 64 times.

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Abstract

The invention relates to a method for preparing Duck hepatitis virus I type indirect blood coagulation diagnosing antigen, in particular to a method for preparing indirect blood coagulation diagnosing antigen by Duck hepatitis virus I type antigen sensibilization double hydroformylation mutton red cell. The Duck hepatitis virus I type indirect blood coagulation diagnosing antigen produced by the method has long storage time up to half a year; moreover, the diagnosing time is shortened and only half an hour is needed, and the use is convenient and simple; therefore, the method is suitable to be popularized and applied to the practical production.

Description

technical field [0001] The invention relates to a preparation method of duck hepatitis type I virus indirect hemagglutination diagnostic antigen, in particular to a method for preparing indirect hemagglutination diagnosis antigen by sensitizing dialdehyded sheep red blood cells with duck hepatitis type I virus antigen. Background technique [0002] Duck viral hepatitis, also known as duck hepatitis, commonly known as "back neck disease", mainly occurs in ducklings under three weeks old. It is a rapidly spreading and highly fatal infectious disease. The disease can be caused by three different pathogens, respectively. Known as DHV-I, DHV-II and DHV-III types; there is no cross-protection between the 3 serotypes. Wherein DHV-I mainly invades ducklings within 4 weeks of age. Especially less than 1-week-old ducklings are the most susceptible to infection, and the mortality rate can reach more than 90%, and the duck virus hepatitis it causes is distributed worldwide, which is on...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/08G01N33/569
Inventor 陶海静
Owner 陶海静
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