Method for preparing trivalent inactivated vaccine for preventing mink hemorrhagic pneumonia
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A technique for hemorrhagic pneumonia and inactivated vaccine, which is applied in the field of preparation of trivalent inactivated vaccine
Inactive Publication Date: 2014-07-30
母连志
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[0004]The pathogen of mink hemorrhagic pneumonia—Pseudomonas aeruginosa is an opportunistic pathogen, which has strong resistance and is prone to drug resistance, so vaccination is an important way to prevent the disease Approach; there is no mink hemorrhagic pneumonia vaccine certified by the state in my country, so it is urgent to develop a mink hemorrhagic pneumonia vaccine targeting the popular serotypes in my country
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example 1
[0012] Example 1, preparation of inactivated vaccine
[0013] 1. Strain rejuvenation
[0014] The SD403, SD302 and SD312 strains were respectively treated with 10LD 50 Inoculate the mice. After the mice die, open the chest cavity under aseptic conditions, cut a small hole in the lung, dip it with an inoculation loop, and streak on the LB solid medium; culture at 37°C for 16 hours, pick a single colony in 5ml In PYG medium, culture at 37°C and 220 rpm for 12 hours, identify by PCR, and preserve the strains. Add 1ml of bacterial solution into 1000ml of PYG medium, incubate at 37°C and 220 rpm for 12h, count viable bacteria, and prepare seeds.
[0015] 2. Inactivated Vaccine Preparation
[0016] (1) Preparation and inspection of bacterial solution
[0017] The SD403, SD302, and SD312 secondary seed liquids that passed the inspection and viable count were inoculated at a ratio of 3% in a fermenter equipped with a PYG liquid medium, ventilated, and incubated at 37°C for 14 hour...
example 2
[0025] Example 2. Inspection of finished inactivated vaccines
[0026] 1 security check
[0027] The inspection items (sterility inspection, safety inspection, formaldehyde residue and thimerosal residue inspection, physical properties) are all carried out in accordance with the "Manufacturing and Inspection Regulations for Veterinary Biological Products" (hereinafter referred to as the "Regulations").
[0028] 1.1 Sterility test
[0029] Refer to the "Rules" for operation; after the sterility test, the three batches of vaccines 201301, 201302, and 201303 all passed the sterility test.
[0030] 1.2 Physical shape inspection
[0031] Observe the color, shape, etc. under the state of standing and shaking respectively; after shaking well, it becomes a uniform brown-yellow suspension; after standing still, the upper layer is a light yellow clear liquid, and the lower layer is a grayish-yellow precipitate;
[0032] 1.3 Inspection of formaldehyde residue and thimerosal residue
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Abstract
The invention relates to a method for preparing a trivalent inactivated vaccine for preventing mink hemorrhagic pneumonia. The method is characterized by comprising the following concrete steps: separating serum G pseudomonas aeruginosa SD302, serum C pseudomonas aeruginosa SD312 and serum I pseudomonas aeruginosa SD403 from an ill mink body; storing bacteria by adopting a freeze-drying mode; respectively putting the three serum pseudomonas aeruginosa into PYG culture medium containing 2% fetal calf serum to cultivate at 37 DEG C; then carrying out enlarge cultivation, and carrying out inactivated treatment on a bacterium solution; diluting the inactivated bacterium solution until the concentration is 8.0*10<9> CFU / mL by using normal saline according to the bacteria content; mixing the three diluted inactivated bacterium solutions with 40% aluminum glue solution according to the volume ratio of 1:1:1:1; adding thiomersalate of which the final concentration is 0.004%, and mixing to obtain the trivalent inactivated vaccine. The method is safe, multi-titer, good in immunogenicity, good in specificity and the like, and is moderate in price, and applicable to substratum or clinical prevention of the mink hemorrhagic pneumonia.
Description
technical field [0001] The invention relates to a preparation method of a trivalent inactivated vaccine for preventing mink hemorrhagic pneumonia. The vaccine belongs to a new animal vaccine and is mainly used to prevent mink hemorrhagic pneumonia caused by Pseudomonas aeruginosa; it is applied in the field of animal husbandry and veterinary medicine . technical background [0002] Mink hemorrhagic pneumonia is an acute contact infectious disease caused by Pseudomonas aeruginosa (also known as Pseudomonas aeruginosa), which is characterized by hemorrhagic pneumonia and sepsis. Since the disease was first discovered in China in 1983, it has been reported successively in various places. Especially since the new century, mink hemorrhagic pneumonia has shown endemic outbreaks in many parts of our country. Together with mink parvovirus and canine distemper, it is called three major infectious diseases that endanger the development of mink breeding industry. [0003] Pseudomonas...
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