Varicella-zoster virus gE antigen and application thereof in detection of anti-varicella-zoster virus antibody

A technology of varicella zoster and virus, applied in the direction of virus/phage, virus, virus peptide, etc., can solve problems such as poor sensitivity

Active Publication Date: 2017-01-04
北京市华信行生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ELISA kits currently available on the market are coated with whole virus antigens, so the sensitivity is poor

Method used

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  • Varicella-zoster virus gE antigen and application thereof in detection of anti-varicella-zoster virus antibody
  • Varicella-zoster virus gE antigen and application thereof in detection of anti-varicella-zoster virus antibody
  • Varicella-zoster virus gE antigen and application thereof in detection of anti-varicella-zoster virus antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Selection of Candidate Antigen Neutralizing Epitopes

[0023] The present inventor carries out antigenicity prediction to varicella zoster virus gE sequence by bioinformatics software, the result is as follows figure 1 As shown, there are 3 main antigenic segments, the sequences of which are shown in SEQ ID NO.1, 2 or 3, respectively.

Embodiment 2

[0024] Example 2 Cloning, expression and activity identification of two gE antigens of varicella-zoster virus

[0025] 1. Design and synthesis of gE antigen gene

[0026] In order to facilitate the efficient expression of the varicella-zoster virus gE gene in E.coli, based on the amino acid sequences of the three varicella-zoster virus gE antigens determined in Example 1, the biased codons of Escherichia coli were selected to design gE Gene, and commissioned Shanghai Yingwei Jieji Biotechnology Service Co., Ltd. to synthesize the gene of the above sequence.

[0027] 2. Construction and transformation of gE antigen expression plasmid

[0028] In order to facilitate the cloning of the gene into the expression vector, two restriction sites Xho I and Xba I were introduced at both ends of the tether to be suitable for the expression vector pBVIL1 (see Chinese patent ZL00100695.9: expression vector pBVIL1 and its construction method and use). Inserted into the vector pBVIL1 after...

Embodiment 3

[0044] Example 3 VZV Antibody Detection

[0045] Use gE antigen 1 (SEQ ID NO.1) and antigen 2 (SEQ ID NO.2) to coat the enzyme-linked plate, and use the indirect ELISA method to use the antibody of goat anti-human IgG to detect the vaccine-induced varicella-zoster virus in the serum Detect with antibody, measurement result and analysis result are as shown in Table 3: Antigen 1 (SEQ ID NO.1) has 1 routine gE-IgG detection result to be positive in the specimen group before 50 routine varicella zoster vaccinations of antigen 1 (SEQ ID NO.1), 49 routine Negative, positive rate was 2%. In the 100-case vaccine group, 90 cases were positive for gE-IgG, 10 cases were negative, and the positive rate was 90%. Antigen 2 (SEQ ID NO.2) 8 cases of gE-IgG test results were positive in the 50 cases of varicella zoster vaccination group before vaccination, 42 cases were negative, and the positive rate was 16%. In the 100-case vaccine group, 89 cases were positive for gE-IgG, 11 cases were ne...

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Abstract

The invention discloses a varicella-zoster virus gE glycoprotein antigen capable of detecting an anti-varicella-zoster virus antibody, particularly an anti-varicella-zoster virus neutralizing antibody, and also discloses application of the antigen in the preparation of a detection reagent for detecting the anti-varicella-zoster virus neutralizing antibody and a corresponding detection reagent kit.

Description

technical field [0001] The invention relates to the field of immunodetection of antibodies, in particular to the field of immunodetection of anti-varicella zoster virus antibodies, especially neutralizing antibodies. Background technique [0002] Chickenpox is a disease caused by primary infection of varicella-zoster virus (VZV). Chickenpox is a highly contagious disease that spreads worldwide. The susceptible population is mainly children, and it is characterized by generalized herpes often accompanied by febrile symptoms. Varicella is a benign, self-limiting disease in the vast majority of immunocompetent children; however, in immunocompromised persons of various causes, VZV infection can cause serious and even fatal disease. Studies have shown that after natural infection or immunization with live attenuated varicella vaccine, the body can be re-infected or activated by varicella virus without any clinical symptoms, and secondary infection with varicella is rare clinica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/04G01N33/68G01N33/569
CPCC07K14/005C12N2710/16722G01N33/56994G01N33/6854G01N2333/04G01N2469/20
Inventor 张贺秋孙妮娜陈庆全
Owner 北京市华信行生物科技有限公司
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