Plasmodium vivax PvMSP1 recombinant antigenic protein as well as preparation method and application thereof

A technology for recombining antigenic proteins and recombinant proteins, which can be used in botany equipment and methods, biochemical equipment and methods, chemical instruments and methods, etc., and can solve the problems of missed detection, malaria parasite infection rate and infection degree, etc.

Active Publication Date: 2014-02-12
中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, malaria in most endemic areas in China is in the elimination stage, and the infection rate and degree of Pla

Method used

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  • Plasmodium vivax PvMSP1 recombinant antigenic protein as well as preparation method and application thereof
  • Plasmodium vivax PvMSP1 recombinant antigenic protein as well as preparation method and application thereof
  • Plasmodium vivax PvMSP1 recombinant antigenic protein as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031]Example 1 Preparation of Plasmodium vivax PvMSP1 Recombinant Antigen Protein

[0032] 1 material

[0033] 1.1 Genomic DNA of Plasmodium vivax

[0034] Serum red blood cells from patients infected with Plasmodium vivax were collected from malaria-endemic areas in Yunnan Province.

[0035] 1.2 Host strains Escherichia coli DH5α, BL21(DE3), and plasmid pET-28a(+) were from Tiangen Biochemical Technology Co., Ltd. and Novagen respectively.

[0036] 1.3 Main reagents and tool enzymes

[0037]

[0038] 2 methods

[0039] 2.1 Amplification and cloning of Plasmodium vivax PvMSP1 gene

[0040] 2.1.1 PCR amplification of PvMSP1 gene

[0041] According to the reported sequence of the PvMSP1 gene (PVX_099980), a pair of specific primers were designed using the primer design software of Shanghai Yingjun Biotechnology Co., Ltd. The uppercase letters in the specific primers were 15 bases homologous to the end of the enzyme-cut vector, Wherein the single underline indicates the...

Embodiment 2

[0078] Example 2 Effect Experiment of Plasmodium vivax PvMSP1 Recombinant Antigen Protein Detection of Human Serum Antibody

[0079] 1 material

[0080] 1.1 Serum samples from vivax malaria patients and normal subjects were collected from malaria endemic areas in Yunnan Province.

[0081] 1.2 Goat anti-human whole molecule IgG, from SIGMA company.

[0082] 2 methods

[0083] A number of patient sera infected by Plasmodium vivax and normal human sera were mixed in equal amounts to prepare positive and negative serum controls. Specific steps are as follows:

[0084] 1) Dilute the antigen with coating solution to a final concentration of 4ng / μl, 100μl / well, incubate at 37°C for 2h, wash with PBS-T 3 times, 3min each time;

[0085] 2) Use 1% BSA (diluted in PBS), 100 μl / well, incubate at 37°C for 1 hour, wash with PBS-T 3 times, 3 minutes each time;

[0086] 3) Dilute the primary antibody (plasma vivax patient serum and normal human serum) with PBS-T (1:100), 100 μl / well, incub...

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Abstract

The invention discloses a plasmodium vivax PvMSP1 recombinant antigenic protein, which is protein of which the amino acid sequence is shown in SEQ ID NO:1 which has glycophosphatidylinositol (GPI) anchor and epidermal growth factor-like (EGF-like) structure domain. Furthermore, the invention also discloses a preparation method of the recombinant antigenic protein, which comprises the steps of amplifying a plasmodium vivax PvMSP1 gene sequence, constructing and identifying recombinant plasmid, inducibly expressing and purifying recombinant protein and the like. Experiments prove that the PvMSP1 recombinant antigenic protein has the advantages of high sensitivity, strong specificity and the like to assay of the serum antibody of a plasmodium vivax infected patient, and has a wide application prospect in the aspect of plasmodium vivax epidemiological investigation.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a recombinant protein, in particular to a Plasmodium vivax PvMSP1 recombinant antigen protein for detecting serum antibodies of patients infected with Plasmodium vivax; in addition, the invention also relates to the preparation of the above-mentioned recombinant antigen protein Methods and uses. Background technique [0002] Malaria is an important parasitic disease that seriously endangers human health and affects social and economic development. Globally, about 1 million people die from malaria parasite infection every year. Malaria (mainly P. vivax) is also one of the important infectious diseases in my country. Although the incidence of malaria has been controlled at a low level, natural and social factors such as global warming and the increase in the flow of people and logistics brought about by the globalization of economic development Under the influence o...

Claims

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Application Information

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IPC IPC(8): C07K14/445C12N15/30C12N15/11C12N15/70G01N33/68G01N33/569C12R1/90
CPCC07K14/445C12N15/70G01N33/56905G01N2333/445Y02A50/30
Inventor 陈军虎樊艳婷陈绅波王越张颋鞠川徐斌胡薇
Owner 中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心
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