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Avian influenza virus H9 subtype positive blood serum and negative blood serum standard substances and preparation methods thereof

A bird flu virus and standard material technology, applied in antiviral immunoglobulin, immunoglobulin from serum, biological testing, etc., can solve problems such as no technical specifications to follow

Inactive Publication Date: 2011-04-13
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 positive blood serum and negative blood serum standard substances and preparation methods thereof
  • Avian influenza virus H9 subtype positive blood serum and negative blood serum standard substances and preparation methods thereof
  • Avian influenza virus H9 subtype positive blood serum and negative blood serum standard substances and preparation methods thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0121] Production of positive serum and inspection of semi-finished products

[0122] 1. Positive serum production

[0123] (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.

[0124] (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 ≥ 8log2 are used for standby.

[0125] (3) Separation of serum Select SPF chickens with HI antibody titer ≥ 8log2 Use a 20ml sterile syringe to collect blood from the heart in ...

Embodiment 2

[0133] Production of weak positive serum and inspection of semi-finished products

[0134] 1. Weak positive serum production

[0135] (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.

[0136] (2) Blood test: 14 to 21 days after immunization, the serum was collected by subwing vein blood collection, 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. SPF chicken with a price of 5log2 is reserved.

[0137] (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 t...

Embodiment 3

[0145] Manufacture of negative serum and inspection of semi-finished products

[0146] 1. Negative Serum Production

[0147] 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.

[0148] 2. Inspection of negative serum semi-finished products

[0149](1) Sterility test Take samples of the mixed serum and test according to "Chinese Veterinary Pharmacopoeia", and there should be no growth of bacteria or mold.

[0150] (2) Titer determination The HI antibody titer should be determined according to the appendix, and it should be negative.

[0151] (3) Specificity Under the same ...

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Abstract

The invention relates to avian influenza virus H9 subtype positive blood serum and negative blood serum standard substances and preparation methods thereof. The standard substance positive blood serum is prepared by using young or adult SPF (specific pathogen free) chicken, carrying out intramuscular injection of an avian influenza virus H9 subtype oil emulsion inactivated vaccine, then collecting blood serum of the SPF chicken, carrying out semi-finished product inspection, adding an appropriate stabilizer and then freeze-drying; the negative blood serum is prepared by collecting the blood of the SPF chicken, separating out the blood serum, carrying out the semi-finished product inspection and freeze-drying; and the preparation is further carried out by a series of technological processes of finished product inspection, homogeneity test, stability test, calibration of the standard substances, value setting and the like. The standard substances are basic guarantee for accurate judgment of avian influenza virus H9 subtype, immunologic surveillance and correct evaluation of immune effect of vaccines of the standard substances, and can be used for improving the prevention and control level of avian influenza. The standard substances constitute the basic guarantee for diagnosis of the avian influenza virus H9 subtype and evaluation and quality control of the related product inspection work.

Description

technical field [0001] The invention relates to a standard substance of avian influenza virus H9 subtype positive serum and negative serum 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 BP1993 edition incl...

Claims

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Application Information

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IPC IPC(8): G01N33/96G01N33/531C07K16/10C07K16/06
Inventor 孙建宏刘景利胡井雷张从禄戴志红韩正博徐姗姗田国彬
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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