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A kind of recombinant human parvovirus B19 protein and its application

A human parvovirus and parvovirus technology, applied in the field of vaccine development, diagnostic reagents, and genetic engineering, can solve the problems of difficulty in antigen source, difficulty in popularization and promotion, etc.

Active Publication Date: 2016-01-13
北京英诺特生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Blood-derived antigens were initially used to detect B19 virus antibodies, but the source of the antigens is difficult, and it is difficult to popularize and promote

Method used

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  • A kind of recombinant human parvovirus B19 protein and its application
  • A kind of recombinant human parvovirus B19 protein and its application
  • A kind of recombinant human parvovirus B19 protein and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] The preparation of embodiment 1 human herpes simplex virus protein

[0062] 1.1 Human herpes simplex virus protein epitope screening and target gene cloning

[0063] The dominant epitopes of the type-specific proteins VP1 and VP2 of the human parvovirus B19 were screened out by computer analysis of the entire amino acid sequence of the human parvovirus B19.

[0064] According to the results of antigenic epitope screening, the polypeptide regions with concentrated 9 epitopes with strong antigenic reaction were selected, and the -SSG- linking arm was used in the middle to list, and the fusion antigen polypeptide expression recombinant sequence was constructed. Design PCR primers P1-P18. Primers P1 and P2 are used to amplify fragments P1P2, P3 and P4 are used to amplify fragments P3P4, P5 and P6 are used to amplify fragments P5P6, P7 and P8 are used to amplify fragments P7P8, P9 and P10 are used to amplify fragments P9P10, P11 and P12 were used to amplify fragment P11P12...

Embodiment 2

[0102] Example 2 Preparation and performance test of human parvovirus B19 type IgM antibody colloidal gold detection kit

[0103] 2.1 Preparation of colloidal gold detection kit for human parvovirus B19 IgM antibody

[0104] The recombinant B19 protein prepared above was used as the labeled antigen of the kit, and the B19 IgM antibody was determined by the colloidal gold method. The kit was developed and used as follows:

[0105] (1) Principle of the kit

[0106] According to the principle of immunocapture method, the invention uses anti-human IgM antibody to coat nitrocellulose membrane, and colloidal gold marks gene engineering recombined parvovirus B19 antigen as a tracer. When using, add the serum to be tested. If the sample contains anti-B19 specific IgM antibody, it can combine with the anti-human IgM antibody on the membrane surface to form a complex, and the complex will combine with colloidal gold-labeled B19 antigen to present a purple band .

[0107] (2) Kit per...

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Abstract

The invention provides a recombinant human parvovirus B19 protein and application of the recombinant human parvovirus B19 protein in preparation of a detection kit. Compared with similar kits on market, the detection kit prepared by adopting the recombinant human parvovirus B19 has the advantages of strong specificity and high sensitivity, and need of clinical diagnosis pm human parvovirus B19 infection is met.

Description

technical field [0001] The invention relates to the fields of genetic engineering, vaccine development, diagnostic reagents and the like, in particular to a human parvovirus B19 protein and its application. Background technique [0002] In 1975, Cossat et al. found spherical virus particles with a diameter of 20-25nm from an asymptomatic healthy blood donor serum sample, which was designated as B19 virus. According to genetic and biochemical analysis, the virus belongs to the genus Parvovirus, called Human Parvovirus B19 (Human Parvovirus B19, hereinafter referred to as B19 virus). B19 virus is currently the only virus that can cause human disease in the parvovirus family except human bocavirus (HBoV) and the new parvovirus PARV4 (Humanparvovirus4). It may be the causative factor of many diseases, infection of children can cause erythema infectious, infection of adults can cause polyarthropathy syndrome, and for some patients with immune diseases and blood diseases, B19 inf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/35C12N15/70C12N1/21C07K14/015G01N33/68G01N33/569C12R1/19
Inventor 张晓刚张秀杰华艳
Owner 北京英诺特生物技术股份有限公司
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