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Chikungunya virus specific detection antigen and application thereof

A virus and auxiliary detection technology, applied in the direction of applications, viruses, virus peptides, etc.

Active Publication Date: 2016-08-31
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, serological methods mainly use inactivated virus cultures as detection antigens for specific antibody detection. In vitro expression of antigens can simplify the antigen preparation process and improve detection specificity. At present, there is no convenient commercial kit available in China.

Method used

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  • Chikungunya virus specific detection antigen and application thereof
  • Chikungunya virus specific detection antigen and application thereof
  • Chikungunya virus specific detection antigen and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Preparation of Chikungunya Virus Specific Detection Antigen

[0053] 1. Chikungunya virus recombinant protein sequence information

[0054] According to the structural characteristics of the E protein of Chikungunya virus, E1 and E2 were divided into different antigen fragments for segmented expression, and specific amplification primers were designed. The fragment length, primer sequence and restriction site information are shown in Table 1.

[0055] Table 1 Fragment length, primer sequence and restriction site information of 3 different antigen fragments

[0056]

[0057] Note: In the table, the underlined part in the upstream primer is the recognition sequence of the restriction site BamHI; the underlined part in the downstream primer is the recognition sequence of the restriction site HindIII.

[0058] 2. Construction of recombinant expression vector

[0059] Use Invitrogen's RNA extraction kit to extract the RNA of Chikungunya virus S27 strain, and use TaKaRa's on...

Embodiment 2

[0075] Example 2. Effect detection of specific detection antigen of Chikungunya virus

[0076] Two different Chikungunya virus E protein fragments (E2-1 and E1-1) prepared in Example 1 were used as detection antigens, and two clinical Japanese encephalitis patients (JEV) infected serum and two forests were collected. Encephalitis (TBEV) patient infection serum samples, 2 dengue (DV) patient infection serum samples, 5 normal human serum samples, 12 chikungunya virus (CHIK) infection serum (the above sera are all clinically confirmed patient sera , Normal human serum was from PLA 307 Hospital, Japanese encephalitis patient serum and forest encephalitis human serum were from Xinjiang Military Region CDC, chikungunya patient serum and dengue patient serum were from Guangzhou Eighth People’s Hospital), and ELISA was used. To evaluate the detection specificity and sensitivity of each recombinant antigen. The steps are as follows:

[0077] (1) The two Chikungunya virus antigens E2-1 and...

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Abstract

The invention discloses a Chikungunya virus specific detection antigen and an application thereof. The Chikungunya virus specific detection antigen provided by the invention is a polypeptide shown as any of (a1), (a2) and (a3), wherein (a1) is a polypeptide shown as a sequence 1 or a 166-378 site of the sequence 1; (a2)is a polypeptide shown as a sequence 2 or a 166-392 site of the sequence 2; and (a3) is a polypeptide which is obtained by performing substitution and / or deletion and / or addition of one or more amino acid residues on an amino acid sequence of the polypeptide defined by any of (a1) and (a2)and has the same function as the polypeptide defined by any of (a1) and (a2). The Chikungunya virus specific detection antigen has relatively high specificity and sensitivity in detection of Chikungunya virus serum infection.

Description

Technical field [0001] The invention belongs to the field of virus detection, and relates to a chikungunya virus specific detection antigen and application thereof. Background technique [0002] Chikungunya disease is a disease caused by chikungunya virus in humans and animals. The patient presented with a sudden onset of high fever and severe joint pain, causing the body to bend and curl up with a skin rash. The disease is mainly transmitted through mosquito bites. In 2010, the disease caused an outbreak in Guangdong, my country. [0003] This virus belongs to the genus Alphavirus of the Epiviridae family. The virus has two structural proteins, membrane protein E and core protein C. The E protein has E1, E2 and E3. E3 is the outer membrane convex that connects E1 and E2 together to form the virus particle. Up. [0004] For the clinical diagnosis of the disease, epidemiological investigations can provide important clues, and suspected cases can be determined based on the patient's ...

Claims

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Application Information

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IPC IPC(8): C07K14/18C12N15/40G01N33/68G01N33/569
CPCC07K14/005C12N2770/36122G01N33/56983G01N33/68G01N2469/20Y02A50/30
Inventor 康晓平杨银辉吴晓燕李裕昌姜涛李靖户义冉鑫贾佳
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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