Cryptosporidium parvum divalent protein vaccine and preparation method thereof

A Cryptosporidium parvum and protein vaccine technology, which is applied in the field of infectious disease vaccine preparation, can solve the problem of no cryptosporidium killing effect, and achieve good prevention of Cryptosporidium parvum in animals, enhanced immune protection effect, and strong immune response The effect of the reaction

Inactive Publication Date: 2010-03-03
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although after many years of research, there is no effective prevention and treatment drug and preventive method so far. More than 200 kinds of drugs have been screened, including all antibiotics, anticoccidiostats and sulfonamides for the treatment of the disease, but there is no specific drug that can cure the disease. play a role in killing Cryptosporidium

Method used

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  • Cryptosporidium parvum divalent protein vaccine and preparation method thereof
  • Cryptosporidium parvum divalent protein vaccine and preparation method thereof
  • Cryptosporidium parvum divalent protein vaccine and preparation method thereof

Examples

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Comparison scheme
Effect test

Embodiment 1

[0017] Preparation of bivalent protein vaccine against Cryptosporidium parvum

[0018] The present invention takes the immune-related protein genes CP12 and CP21 of Cryptosporidium parvum as an example to construct the recombinant prokaryotic expression vector.

[0019] The preparation steps of Cryptosporidium parvum bivalent protein vaccine:

[0020] Refer to the open reading frames of CP12 and CP21 gene sequences and the physical map of prokaryotic expression vector pET28a to design primers and introduce restriction sites.

[0021] CP12 upstream primer:

[0022] QF1: 5'-CT GGATCC ATGTCAGATGCATCAATA-3'; where the 5' end contains a BamHI site;

[0023] Downstream primer QR1: 5'-GGGGC GAATTC TATTTGTTCATTCATCTG-3'; 5' end contains an EcoRI site.

[0024] CP21 upstream primer:

[0025] QF2: 5'-CC GAATTC GGTGGCGGTGGCTCGATGTCTAAAAAGAGCAT-3', the 5' end contains EcoRI site and Linker sequence;

[0026] Downstream primer QR2: 5'-CTGGCT CTCGAG CTATTCATCTGTTTGAAC-3'; 5' en...

Embodiment 2

[0029] Uses of Cryptosporidium parvum Bivalent Protein Vaccine

[0030] 1. Application of Cryptosporidium parvum bivalent protein vaccine

[0031] Divide 60 4-6 weeks old, 18-22g female clean BALB / c mice into 3 groups, 20 mice in each group. The experimental group was intraperitoneally injected with recombinant protein, and immunized three times with an interval of 2 weeks between each time. For the first immunization, 0.1 mL antigen solution (containing 20 μg recombinant protein) was emulsified with an equal volume of Freund’s complete adjuvant and injected intraperitoneally; for the second and third immunizations, 0.1 mL antigen solution (containing 20 μg recombinant protein) was mixed with an equal volume of The incomplete adjuvant was emulsified and injected intraperitoneally. The adjuvant control group (Negative control B) was intraperitoneally injected with the same amount of Freund's adjuvant as the test group, the first injection of Freund's complete adjuvant, the sec...

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Abstract

The invention provides a cryptosporidium parvum divalent protein vaccine, which is characterized in that prokaryotic expression vector pET28a is taken as a carrier, and two cryptosporidium parvum protective antigen genes are inserted in series to the multiple cloning sites of the prokaryotic expression pET28a, thus obtaining the cryptosporidium parvum divalent protein vaccine. The vaccine selectstwo advantageous protective antigens, combines the advantages of the two antigens, and can induce stronger functions of cellular immunity and humoral immunity, thus achieving the purposes of preventing cryptosporidium parvum disease of animal better.

Description

technical field [0001] The invention provides a cryptosporidium parvum bivalent protein vaccine, which is used for the prevention and treatment of cryptosporidiosis, and also provides a preparation method of the vaccine, which belongs to the technical field of infectious disease vaccine preparation. Background technique [0002] Cryptosporidium (Cryptosporidium) is a zoonotic parasitic protozoan, mainly parasitic in the epithelial cells of the gastrointestinal tract of humans and other mammals, the respiratory tract of birds, etc., and the disease caused by it is called cryptosporidiosis . Cryptosporidiosis is widely distributed, with a high infection rate in humans and animals, and serious harm. Clinically, severe watery diarrhea is the main feature. The infection rate of Cryptosporidium in AIDS patients can reach 10% to 50%, and it is one of the main causes of death in AIDS patients. Among all Cryptosporidium species, Cryptosporidium parvum is the most threatening to hum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/015A61P33/02
CPCY02A50/30
Inventor 宫鹏涛李建华张西臣于钦磊李淑红杨举
Owner JILIN UNIV
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