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Positive-serum and negative-serum standard substance of avian influenza virus H5N1 subtype Re-5 strain and preparation method thereof

An avian influenza virus, standard material technology, applied in the direction of anti-animal/human immunoglobulin, etc., can solve the problem of no technical specifications to follow

Inactive Publication Date: 2011-05-11
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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  • Positive-serum and negative-serum standard substance of avian influenza virus H5N1 subtype Re-5 strain and preparation method thereof
  • Positive-serum and negative-serum standard substance of avian influenza virus H5N1 subtype Re-5 strain and preparation method thereof
  • Positive-serum and negative-serum standard substance of avian influenza virus H5N1 subtype Re-5 strain and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] Production of positive serum and inspection of semi-finished products

[0127] 1. Positive serum production

[0128] (1) For immunization, select SPF chickens aged 3 to 4 months and raise them in a negative pressure isolator, and use the prepared avian influenza inactivated vaccine to vaccinate twice by chest intramuscular injection. After 21 days, the second inoculation was carried out, the dose was 1.5ml / only, and the injection was divided into points.

[0129] (2) Blood test: 28 days after the second immunization, the serum was collected by means of subwing vein blood collection. At the same time, the chicken and serum were numbered correspondingly, and the HI antibody titer was determined respectively (according to the "Appendix"), and the HI antibody was selected. SPF chickens with a titer ≥ 9log2 are used for standby.

[0130] (3) Separation of serum Select SPF chickens with HI antibody titer ≥ 9log2 Use a 20ml sterile syringe to collect blood from the heart in ...

Embodiment 2

[0138] Production of weak positive serum and inspection of semi-finished products

[0139] 1. Weak positive serum production

[0140] (1) Immunization SPF chickens aged 3 to 4 months were selected and raised in a negative pressure isolator, and the prepared inactivated avian influenza vaccine was vaccinated by chest intramuscular injection, and the inoculation dose was 0.5ml / bird.

[0141] (2) For blood test, 14 to 21 days after immunization, the serum was collected by the subwing vein blood collection method, and the chicken and serum were correspondingly numbered at the same time, and the HI antibody titer was determined respectively (according to the appendix), and the HI antibody titer was selected as 5log2 SPF chicken for backup.

[0142] (3) Separation of serum Select SPF chickens with HI antibody titer 5log2 and use a 20ml sterile syringe to collect blood from the heart in a negative pressure isolator. After the blood is collected, place it at 37°C for 4 hours, and the...

Embodiment 3

[0150] Manufacture of negative serum and inspection of semi-finished products

[0151] 1. Negative Serum Production

[0152] The young SPF chickens were raised in the negative pressure isolator in the biosafety animal laboratory, and the heart blood was collected with a sterile syringe in the negative pressure isolator in the biosafety animal laboratory. After the blood was collected, it was placed at 37°C for 4h. Then put it in the refrigerator at 4°C overnight, extract the serum in the ultra-clean workbench in the sterile room, and then mix it in the same sterilized container.

[0153] 2. Inspection of negative serum semi-finished products

[0154] (1) Bacteria test Take samples of the mixed serum and test according to "Chinese Veterinary Pharmacopoeia", and there should be no bacteria or mold growth.

[0155] (2) Titer determination The HI antibody titer should be determined according to the appendix, and it should be negative.

[0156] (3) Specificity Under the same con...

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Abstract

The invention relates to a positive-serum and negative-serum standard substance of avian influenza virus H5N1 subtype Re-5 strain and a preparation method thereof. The preparation method comprises the following steps of: preparing a standard substance positive serum (strongly positive serum and weakly positive serum): intramuscularly injecting an avian influenza virus H5N1 subtype Re-5 strain oil-emulsion inactivated vaccine in a young or adult SPF (Specific Pathogen Free) chicken, and then collecting the serum of the SPF chicken and carrying out semi-finished product inspection, adding a proper stabilizing agent and then freeze-drying; preparing negative serum: taking the blood of the SPF chicken, separating serum, carrying out semi-finished product inspection, and freeze-drying; and carrying out a series of technical processes, such as finished-product inspection, uniformity inspection, stability inspection, calibration and value determination of the standard substance and the like to obtain the positive-serum and negative-serum standard substance. The standard substance is the fundamental guarantee for the accurate diagnosis of avian influenza virus H5N1 subtype, the immune monitoring of a H5N1 subtype Re-5 strain and the accurate evaluation on the vaccine immune effect of the H5N1 subtype Re-5 strain, thereby improving the prevention and control level of avian influenza. The standard substance is the basic guarantee for the diagnosis of avian influenza virus H5N1 subtype and the evaluation and the quality control of the inspection working of related products.

Description

technical field [0001] The invention relates to a positive serum and a negative serum standard substance of an avian influenza virus H5N1 subtype Re-5 strain and a preparation method thereof, 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 ...

Claims

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Application Information

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IPC IPC(8): C07K16/18
Inventor 孙建宏刘景利田国彬胡井雷杨帆徐姗姗张从禄韩正博
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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