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Japanese blood fluke nucleic acid vaccine

A schistosomiasis nucleic acid and schistosomiasis technology, which is applied in the field of nucleic acid vaccines against schistosomiasis, can solve the problems of long preparation process, high cost of manpower and material resources, time-consuming and the like

Inactive Publication Date: 2004-07-14
华中科技大学同济医学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Live attenuated schistosomiasis cercariae vaccines require specific equipment, which is inconvenient to carry, limited sources of quantity, virulence recovery of schistosomiasis cercariae, that is, safety problems; the preparation of genetically engineered protein vaccines is cumbersome, the preparation process is long, time-consuming, laborious, manpower and material resources Larger, limited promotion and application

Method used

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  • Japanese blood fluke nucleic acid vaccine
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  • Japanese blood fluke nucleic acid vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0007] Example 1 Preparation of monovalent DNA vaccines (pCD-Sj26, pCD-Sj32, pCD-Sj23 and pBK-Sj26)

[0008] 1. Design primers according to the nucleotide sequences of Sj23, Sj26 and Sj32. The nucleotide sequences of the three pairs of primers are respectively listed in sequences 4 to 9 in the sequence table, and were synthesized by Shanghai Sangon Biotechnology Co., Ltd.

[0009] See sequence 4 for the primer at the 5' end of Sj23; see sequence 5 for the primer at the 3' end of Sj23.

[0010] See sequence 6 for the primer at the 5' end of Sj26; see sequence 7 for the primer at the 3' end of Sj26.

[0011] See sequence 8 for the primer at the 5' end of Sj32; see sequence 9 for the primer at the 3' end of Sj32.

[0012] 2. Extract the total RNA of Schistosoma japonicum, and carry out reverse transcription PCR, i.e. RT-PCR, with the above three pairs of primers respectively, the method is as follows:

[0013] Weigh 100mg adult worms, add 1ml TRI 20L The reagent (GIBCO) was ho...

Embodiment 2

[0094] Example 2 Preparation of multivalent DNA vaccine (pBK-Sj26-Sj23)

[0095] In order to improve the protection against schistosomiasis infection induced by Schistosoma japonicum DNA vaccine, the present invention provides a mixed DNA vaccine (Cocktail vaccine, i.e. cocktail vaccine) and a multivalent DNA vaccine, that is, two candidate vaccine molecules are connected by a base linker of a specific polypeptide .

[0096] 1. The base linker sequence of the polypeptide:

[0097] According to the 26KDa (Sj26) and 23KDa (Sj32) sequences of Schistosoma japonicum and the multiple cloning site of the eukaryotic expression vector pBK, 2 pairs of primers were designed respectively, at the 3' end of Sj26, that is, one of the primers 2 of Sj26 was designed with the Sj23 5' The nucleotide sequence of the enzyme cleavage site XbaI with the same end and the polypeptide linker G-G-S with better mobility, the primer sequence is as follows:

[0098] Sj26: Primer 1: 5'-TAAGGATCCCATGTCCCCC...

Embodiment 3

[0106] Embodiment 3 Effect evaluation experiment of the present invention

[0107] Experiments have proved that the schistosomiasis DNA vaccine of the present invention can induce better protection. Because schistosomiasis does not reproduce in the body, one is eliminated and one is reduced. Therefore, the schistosomiasis vaccine is different from other infectious disease vaccines. WHO points out that the vaccine induces the protection of the host against schistosomiasis. If the force can reach 50%, it can be considered that the degree of protection induced is relatively high. The observation indicators of this experiment are:

[0108] 1. Insect reduction rate:

[0109] Group Number of challenged cercariae After challenge infection Reduction rate (%) p-value

[0110] Number of adults seized

[0111] Blank control group 40 30.60±5.32 -- --

[0112] DNA vaccine pCD-Sj26 group 40 19.22±4.12 37.19 <0.01

[0113] DNA vaccine pCD-Sj32 group 4...

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Abstract

The Japanese blood fluke nucleic acid vaccine consists of eukaryotic expression vector pCD or pBK and blood fluke antigen genes. The blood fluke antigen genes includes Japanese blood fluke antigen genes Sj-23, Sj-26 and Sj-32 and constitute monovalent nucleic acid vaccines pCD-Sj23, pCD-Sj26, pCD-Sj32 and pBK-Sj26 and multivalent nucleic acid vaccine pBK-Sj26-Sj23. The vaccines have blood fluke reducing rate of 35.4-44.4 % and blood fluke egg reducing rate in liver of 39.6-69.0 %, and can induce effectively the immunological protecting capacity to blood fluke. Once immunization has relatively long immunological effect, and the present invention has no environmental pollution, bad influence, and bad reaction.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a nucleic acid vaccine for zoonotic parasitic diseases, in particular to a nucleic acid vaccine for schistosomiasis. Background technique [0002] Schistosomiasis is a zoonotic parasitic disease that endangers human health and affects national economic development. According to WHO data in 1990, schistosomiasis is distributed in 76 countries and regions. It is estimated that 500-600 million people are threatened, and the number of patients reaches 200 million. There are still 1.5 million patients in my country, and cattle infection is also quite serious, which is a major public health problem in developing countries including China. For a long time, people have pinned their hopes on the research and development of effective vaccines for the effective prevention and treatment of schistosomiasis. The existing schistosomiasis vaccines mainly include live attenuated vaccines and genetically...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K45/00A61P33/12
Inventor 石佑恩李柳哲
Owner 华中科技大学同济医学院
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