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Sendai virus antigen peptide composition and application thereof in detecting Sendai virus infection

A Sendai virus and infection detection technology, applied in the field of biomedicine, can solve problems such as affecting antigen stability, affecting the results of antigen epitope detection, affecting accuracy, etc., to reduce the possibility of cross-reaction, improve specificity and target performance, improve accuracy

Active Publication Date: 2013-06-19
INST OF LAB ANIMAL SCI CHINESE ACAD OF MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the existence of many variables in the process of amplification, culture and isolation and purification of whole virus particles in the laboratory, including the variation of the pathogen itself, the non-specific antigenic components that may be incorporated into each virus production, etc., these factors will affect the antigen stability
Although the whole virus particle guarantees the existence of all antigenic sites, different ways of antigen exposure will also affect the display of antigenic epitopes and finally affect the results of detection work
Moreover, the virus whole particle antigen will produce cross-antigens with other pathogens, which will eventually affect the accuracy of detection.

Method used

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  • Sendai virus antigen peptide composition and application thereof in detecting Sendai virus infection
  • Sendai virus antigen peptide composition and application thereof in detecting Sendai virus infection
  • Sendai virus antigen peptide composition and application thereof in detecting Sendai virus infection

Examples

Experimental program
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Effect test

Embodiment 1

[0200] Determination of the dominant antigenic protein of embodiment 1 Sendai virus

[0201] First, prepare the solution for SDS-PAGE electrophoresis:

[0202] 1.0mol / L Tris (pH6.8) solution: Dissolve 12g Tris in 80mL distilled water, adjust the pH value to 6.8, and dilute to 100mL with distilled water;

[0203] 1.5mol / L Tris (pH8.8) solution: Dissolve 18g Tris in 80mL distilled water, adjust the pH value to 8.8, and dilute to 100mL with distilled water;

[0204] 10% SDS solution: Dissolve 10g of SDS in 80mL of distilled water, adjust the pH value to 7.2, and dilute to 100mL with distilled water;

[0205] 10% ammonium persulfate solution: Dissolve 1g of ammonium persulfate in 10mL of distilled water;

[0206] 5×Tris-glycine electrophoresis buffer: Tris15.1g, glycine 94g, completely dissolved in 800mL distilled water, add SDS5g, dilute to 1L, dilute to 1× before use;

[0207] 30% acrylamide stock solution: Dissolve 1g of methylenebisacrylamide in 80mL of water and heat to di...

Embodiment 2

[0212] Example 2 Screening of Sendai virus dominant antigen polypeptide epitope

[0213] According to the protein sequence of 524 amino acids of NP protein, two polypeptide arrays were made by polypeptide array technology. The array chip is designed in an overlapping manner, calculated on the basis of 14 amino acid polypeptide lengths and 2 amino acid displacements, 264 polypeptides are arranged on each polypeptide array, and a total of 528 polypeptides are arranged on two array chips.

[0214] Configure the required solution:

[0215] Concentration is the Fmoc-amino acid solution of 0.25mol / L;

[0216] Blocking solution I is a dimethylformamide solution containing 2% (v / v) acetic anhydride;

[0217] Blocking solution II is 2% (v / v) of acetic anhydride and 2% (v / v) of N,N-diisopropylethylamine in dimethylformamide;

[0218] The Fmoc-removal solution was a solution in dimethylformamide containing 20% ​​(v / v) piperidine.

[0219] First, synthesize the peptide array:

[0220...

Embodiment 3

[0225] Example 3 Functional Identification of Sendai Virus Antigen Polypeptide Composition

[0226]The 4 polypeptides screened out in Example 2 of the present invention were prepared by chemical synthesis, named as Antigen A, Antigen B, Antigen C, and Antigen D respectively, and were coated into ELISA reaction plates at a concentration of 1 mg / mL, and respectively mixed with Sendai Virus-positive sera react with negative sera, and the results are as follows Figure 4 shown. The results showed that the 4 peptides were less immunogenic than the natural antigen SeV (Sendai virus). However, antigen A, antigen C and antigen D still showed good immunogenicity, and antigen B had poor antigenicity. Wherein, the amino acid sequence of antigen A is shown in SEQ ID NO:1, the amino acid sequence of antigen B is shown in SEQ ID NO:2, and the amino acid sequence of antigen C is shown in SEQ ID NO:3.

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Abstract

The invention relates to the technical field of biomedicine, particularly a Sendai virus antigen peptide composition and application thereof in detecting Sendai virus infection. The Sendai virus antigen peptide provided by the invention comprises polypeptide with the sequence disclosed as SEQ ID NO:1, polypeptide with the sequence disclosed as SEQ ID NO:2 and polypeptide with the sequence disclosed as SEQ ID NO:3, or comprises polypeptide subjected to substitution, deletion or addition of one or more amino acid residues on the sequence disclosed as SEQ ID NO:1, polypeptide subjected to substitution, deletion or addition of one or more amino acid residues on the sequence disclosed as SEQ ID NO:2 and polypeptide subjected to substitution, deletion or addition of one or more amino acid residues on the sequence disclosed as SEQ ID NO:3. The Sendai virus antigen peptide composition provided by the invention has lower molecular weight, lowers the possibility of cross reaction with other molecules, and enhances the detection accuracy.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a Sendai virus antigen peptide composition and its application in the detection of Sendai virus infection. Background technique [0002] Sendai virus (Sandaivirus) belongs to Paramyxoviridae Paramyxoviridae subfamily, Respirovirus, is a single-stranded negative-sense RNA virus, is a common and highly infectious parainfluenza virus in experimental rodents, and is the cause of infection in rodents. The main pathogen of respiratory diseases, rodent experimental animals such as mice, rats, hamsters and guinea pigs are susceptible. Sendai virus has no obvious clinical manifestations in most cases, but it will affect the immune system of experimental animals and cause lethal effects after infection with other bacterial pathogens. It is a pathogen that must be checked for rodent experimental animals and is highly contagious. , easy to spread, and in most cases it is a recessive inf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/45A61K39/155A61P31/14G01N33/68
Inventor 向志光杨志伟佟巍刘先菊李雨函魏强
Owner INST OF LAB ANIMAL SCI CHINESE ACAD OF MEDICAL SCI
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