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Enterovirus 71 capsid protein 1 antigen, preparation method and immunodetection reagent

An enterovirus, capsid protein technology, applied in antiviral agents, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of limited application, inability to prepare in large quantities, low gene expression, etc. dangerous effect

Inactive Publication Date: 2011-11-16
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because the VP1 antigen gene contains a large number of rare codons for Escherichia coli, the gene expression level is low, and it cannot be produced in large quantities, which limits its application in EV71-IgM ELISA detection reagents and vaccines

Method used

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  • Enterovirus 71 capsid protein 1 antigen, preparation method and immunodetection reagent
  • Enterovirus 71 capsid protein 1 antigen, preparation method and immunodetection reagent
  • Enterovirus 71 capsid protein 1 antigen, preparation method and immunodetection reagent

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Embodiment Construction

[0027] 1. Transformation and synthesis of EV71-VP1 gene.

[0028] We used bioinformatics software to reverse-translate the amino acid sequence of the EV71-VP1 antigen into a nucleotide sequence using the dominant codons of E. coli. The modified EV71-VP1 gene sequence is shown in sequence 1 in the sequence table; Domestic gene primer synthesis efficiency, the present invention divides the above-mentioned gene into four sections of E, F, G, and H and synthesizes them respectively. The end of the four gene fragments has a matching sequence of 16 nucleotides. After further splicing into a complete EV71-VP1 gene sequence, each synthetic primer sequence is shown as 2-33 in the sequence table. In order to facilitate the description of the preparation method, the corresponding gene sequence is named, SEQ ID NO: 2 to 5 respectively Named as EF4, EF3, EF2, EF1; SEQ ID NO: 6 to 9 are respectively named ER1, ER2, ER3, ER4; SEQ ID NO: 10 to 13 are respectively named FF4, FF3, FF2, FF1; SEQ...

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Abstract

The invention discloses an escherichia coli optimal codon spliced enterovirus 71 (EV71) capsid protein 1 (VP1) antigen gene sequence. According to the invention, a reconstructed VP1 gene is obtained directly from 32 pieces of gene primer fragments through an annealing extension PCR (Polymerase Chain Reaction) technology, without an EV71 virus template. A reconstructed VP1 gene has a high expression level, can be purified to obtain a corresponding recombinant EV71-VP1 antigen. In addition, the method has higher security than a virus cracking method.

Description

technical field [0001] The invention relates to enterovirus 71 (EV71) capsid protein 1 (VP1) gene modification and recombinant antigen, and also relates to a detection reagent for detecting EV71 antibody. Background technique [0002] Enterovirus 71 (Enterovirus 71, EV71) infection mainly causes hand, foot and mouth disease (HFMD) in patients clinically. The main target of infection is children under 5 years old. The positive rate of serum anti-EV71 antibody level in children under 2 years old is only 0.8%, and the serum antibody level rises and reaches a stable state at about 50% at the age of 5-6 years; gradually decreased with increasing age. [Ooi E E, Phoon M C, Ishak B, et al. Seroepidemiology of human enterovirus 71. Emerg Infect Dis. 2002; 8: 995-997.]. Adults can also be infected and spread within the same family members, but the clinical symptoms are not obvious. This subclinical infection or recessive infection is another important way for the rapid spread of E...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/41C12N15/10G01N33/569A61K39/125A61P31/14C12R1/19C12R1/93
Inventor 修冰水陈堃张贺秋王国华宋晓国
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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