Recombinant human parvovirus B19 protein and application thereof
A technology of human parvovirus and B19, which is applied in the field of vaccine development, diagnostic reagents, and genetic engineering, and can solve the problems of difficult source of antigen, difficult popularization and promotion, etc.
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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
[0103] Example 2 Preparation and performance test of human parvovirus B19 type IgM antibody colloidal gold detection kit
[0104] 2.1 Preparation of colloidal gold detection kit for human parvovirus B19 IgM antibody
[0105] 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:
[0106] (1) Principle of the kit
[0107] 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 .
[0108] (2) Kit per...
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