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Avian influenza virus H9 subtype hemagglutination inhibition antigen standard substance and preparation method thereof

A bird flu virus and standard material technology, applied in the field of bird flu virus H9 subtype hemagglutination inhibitor standard material and preparation field, can solve the problem of no technical specification to follow

Inactive Publication Date: 2011-04-06
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the development of reference materials in my country mainly refers to and follows the "Management Measures for Standard Materials" issued and implemented by the National Bureau of Metrology on July 10, 1987, and the technical specifications for primary standard materials (JJG 1006-94), but this technical specification is applicable to chemical Composition, physical and chemical characteristics and engineering technical characteristics of the development of primary reference materials, there is currently no technical specification for the development of reference materials for biological products, especially reference materials for veterinary biological products

Method used

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  • Avian influenza virus H9 subtype hemagglutination inhibition antigen standard substance and preparation method thereof
  • Avian influenza virus H9 subtype hemagglutination inhibition antigen standard substance and preparation method thereof
  • Avian influenza virus H9 subtype hemagglutination inhibition antigen standard substance and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Antigen manufacturing and inspection of semi-finished products

[0094] 1 Preparation of poisonous seeds for production

[0095](1) Propagation of the virus seeds The virus seeds are diluted serially 10 times with sterilized normal saline, and the dilution degree of 10-4 is taken, and 11-day-old SPF chicken embryos are inoculated in the allantoic cavity, with 0.1ml of each embryo, the pinholes are sealed, and placed at 37 Continue to incubate at ℃ without turning the eggs. After 24 hours of inoculation, the eggs were illuminated once every 8 hours, until 72 hours, the dead chicken embryos were taken out at any time, the air chamber was upright, cooled at 2-8°C, and those that died before 24 hours after inoculation were discarded. Take out the chicken embryos that have been cooled for 8-24 hours, disinfect the air chamber with tincture of iodine, then use aseptic surgery to remove the egg shells in the air chamber, remove the yolk shell membrane, cut the chorioallantoic...

Embodiment 2

[0113] Product testing

[0114] 1 Physical properties White or off-white spongy loose lumps, easy to separate from the bottle wall, dissolve quickly after adding diluent.

[0115] 2 Sterility test Sampling test, there should be no growth of bacteria or mold. The test results are shown in Table 1, and all freeze-dried products were all negative.

[0116] Table 1 Statistical table of sterility testing

[0117]

[0118] 3 Potency determination Samples were randomly selected from different parts during freeze-drying for hemagglutination determination (performed in accordance with Appendix 1), and the determination results are shown in Table 2. The results show that the agglutination value of 1% chicken red blood cells is not lower than 7log2, reaching 9log2.

[0119] Table 2 01 Antigen Titer Test Results

[0120]

[0121] Note: "#" in the table indicates complete agglutination, "++" indicates partial agglutination, and "-" indicates no agglutination.

[0122] 4 The spec...

Embodiment 3

[0131] 1 Valuation of standard substance potency

[0132] (1) Fixed value method

[0133] Erythrocyte agglutination test was carried out by 96-well microplate method.

[0134] (2) fixed value unit

[0135] 6 to 8 laboratories with the same qualifications and confirmed by comparison in advance that they have the same value-setting ability will use the same method to carry out collaborative calibration.

[0136] (3) Fixed value requirements

[0137] 1) Six to eight independent measurements should be made for each standard sample to be tested, divided into two units, and three independent measurements should be made for each unit; the interval between the two units should not be less than 3 days. The 6 data obtained should be tested for abnormal values ​​according to statistical methods. If abnormal values ​​are found, they should be marked and a supplementary measurement should be made. Then report all the results.

[0138] 2) All measuring instruments / measuring instruments...

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Abstract

The invention relates to an avian influenza virus H9 subtype hemagglutination inhibition antigen standard substance and a preparation method thereof. The preparation method comprises the following steps: preparing a virus liquid of avian influenza virus H9 CKSH01 strain virus, and inspecting; inactivating the virus liquid, concentrating, and inspecting the semi-finished product; and freeze-drying, inspecting the finished product, inspecting the uniformity and stability, calibrating the standard substance, fixing the value and the like to obtain the standard substance. The standard substance is the basic guarantee for accurate diagnosis and immunity monitoring of avian influenza virus H9 and the proper evaluation on the vaccine immunity effect, and enhances the level for preventing and controlling avian influenza.

Description

technical field [0001] The invention relates to a bird flu virus H9 subtype hemagglutination inhibiting antigen standard substance and a preparation method, belonging to the field of veterinary biological products. Background technique [0002] The chemical reference substances of the British Pharmacopoeia (BP) were only proposed in 1963. Since the WHO established an international biological standard substance center in the United Kingdom, the United Kingdom has always used international standard substances, while its own national standard substances were established later, after 1970. , the European Pharmacopoeia came out. In addition to citing international standard substances, the UK also used European Pharmacopoeia standard substances, and used its own standard substances in a small amount. Even so, the BP1968 edition and its supplement plus the British Pharmacopoeia (1968) included more than 260, the 1980 edition exceeded 300, and the BP1993 edition included 371. The U...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531C12N7/04
Inventor 孙建宏刘景利胡井雷杨帆王在时韩正博张从禄田国彬
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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