Recombinant human Coxsackie virus group-B proteins and application thereof

A coxsackie virus and histone technology, which is applied in the fields of genetic engineering, vaccine development and diagnostic reagents, can solve the problem of long time-consuming separation and culture, and achieve the effect of meeting the needs of clinical diagnosis, high sensitivity and high specificity

Active Publication Date: 2014-06-04
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

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Problems solved by technology

However, isolation and culture takes a long time, and isolation must be perf

Method used

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  • Recombinant human Coxsackie virus group-B proteins and application thereof
  • Recombinant human Coxsackie virus group-B proteins and application thereof
  • Recombinant human Coxsackie virus group-B proteins and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0048] The preparation of embodiment 1 recombinant human coxsackie virus B group protein

[0049] 1.1 Recombinant human Coxsackievirus B group protein antigen epitope screening and target gene cloning

[0050] By computer analysis of the entire amino acid sequence of Coxsackie virus type B4 (Genbank: AFR79234.1, strain JX308222.1), the dominant antigenic epitopes of the specific proteins of human Coxsackie virus group B, including coxsackie The 273 nucleotides of the N-terminal B histone protein from 1162 to 1434 (fragment 1) and the 345 nucleotides from 2626 to 2970 of the N-terminal (fragment 2). The recombinant DNA sequence encoding the recombinant protein is shown in SEQ ID No.1.

[0051] According to the DNA sequence of the target fragment, that is, the cDNA sequence of the target peptide of COXB and the restriction site on the plasmid, two pairs of PCR primers (P1, P2) and (P3, P4) were designed. Primers P1 and P2 are used to amplify fragment 1, primers P3 and P4 are u...

Embodiment 2

[0070] Example 2 Preparation and Performance Test of Recombinant Human Coxsackievirus Group B IgG Antibody Colloidal Gold Detection Kit

[0071] 2.1 Preparation of Colloidal Gold Detection Kit for Recombinant Human Coxsackievirus Group B IgG Antibody

[0072] The recombinant COXB protein prepared above was used as the labeled antigen of the kit, and the COXB IgG antibody was determined by the colloidal gold method. The kit was developed and used as follows:

[0073] (1) Principle of the kit

[0074] According to the principle of immune capture method, the invention uses anti-human IgG (anti-γ chain) antibody and goat anti-mouse IgG antibody to coat nitrocellulose membrane, and colloidal gold labeled with recombinant human COXB antigen and mouse IgG antibody is used as a tracer. When using, add the serum to be tested. If the sample contains COXB IgG antibody, it can combine with the colloidal gold-labeled COXB antigen to form a complex. A purple-red band appears as a detecti...

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Abstract

The invention provides a recombinant DNA for encoding recombinant human Coxsackie virus group-B proteins, a recombinant human Coxsackie virus group-B protein, a method for preparing the recombinant human Coxsackie virus group-B protein, a kit for detecting human Coxsackie virus group-B infection as well as application of the recombinant human Coxsackie virus group-B protein for detecting the human Coxsackie virus group-B infection. The recombinant DNA has a nucleotide sequence shown as SEQ ID NO:1. Compared with a similar kit in the market, the diagnostic kit prepared by the recombinant human Coxsackie virus group-B protein provided by the invention has the advantages of high specificity, high sensitivity and the like, and the requirement on clinical diagnosis of the human Coxsackie virus group-B infection is well 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 recombinant DNA encoding recombinant human coxsackievirus group B protein, a recombinant human coxsackievirus group B protein and applications thereof. Background technique [0002] Coxsackievirus (Coxsackievirus) is a kind of enterovirus (enteroviruses), divided into two types A and B, is a common virus that infects the human body through the respiratory tract and digestive tract. After infection, people will have cold symptoms such as fever, sneezing, and coughing. Infection during pregnancy can cause non-paralytic poliomyelitis lesions, and cause intrauterine infection and teratogenicity of the fetus. [0003] There are 23 types of coxsackieviruses in group A and 6 types in group B. Various types can be identified through type-specific antigens, neutralization tests, and ELISA methods. [0004] All coxsackieviruses...

Claims

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Application Information

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IPC IPC(8): C12N15/41C07K14/085C12N15/70C12N1/20C12Q1/70C12Q1/68C12R1/19
Inventor 张秀杰华艳张晓刚
Owner 北京英诺特生物技术股份有限公司
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