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Method for detecting type-5, type-7 and type-55 adenoviruses by using mass spectrum multiple reaction monitoring technology

A multiple reaction and technical detection technology, applied in the field of biomedicine, can solve the problems of inability to use early warning adenovirus infection, lack of high-efficiency specific monoclonal antibodies, etc., and achieve the effects of large popularization and application value, easy operation and accurate results.

Pending Publication Date: 2021-07-23
LOGISTICS UNIV OF CAPF
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even though some antigen detection reagents targeting hexon proteins have been developed, they cannot be used for early warning of adenovirus infection due to the lack of efficient and specific monoclonal antibodies and the clinical specimens used to evaluate the specificity and sensitivity of detection reagents

Method used

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  • Method for detecting type-5, type-7 and type-55 adenoviruses by using mass spectrum multiple reaction monitoring technology
  • Method for detecting type-5, type-7 and type-55 adenoviruses by using mass spectrum multiple reaction monitoring technology
  • Method for detecting type-5, type-7 and type-55 adenoviruses by using mass spectrum multiple reaction monitoring technology

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Embodiment 1

[0049] The invention provides a method for detecting type 5, 7, and 55 adenoviruses using mass spectrometry multiple reaction monitoring technology, the specific steps of which include:

[0050] Step (1): Synthesizing adenovirus type 5, type 7, and type 55 hexon protein-specific peptide genes;

[0051] Step (2): Preparation of labeled peptide mixed reagents;

[0052] Step (3): determining the ion peak value of the diagnostic marker;

[0053] Step (4): Carry out the preparation of kit;

[0054] Step (5): Perform sample detection.

[0055] The specific method for synthesizing the hexon protein specific peptides of adenovirus type 5, type 7, and type 55 is as follows: compare the hexon protein sequences of adenovirus type 5, type 7, and type 55 with a bioinformatics database, and select specific The optimal protein sequence, through the theoretical digestion scheme, select 13 landmark peptides for type 5 hexon protein, select 10 landmark peptides for type 7 hexon protein, and ...

Embodiment 2

[0073] Technical route such as figure 1 shown.

[0074] Step (1): Synthesis of adenovirus type 5, type 7, and type 55 hexon protein-specific peptide genes

[0075] The bioinformatics database compared the hexon protein sequences of adenovirus type 5, type 7, and type 55, and selected the best specific protein sequence. Through the theoretical digestion scheme, 13 iconic peptides were selected for the type 5 hexon protein. Select 10 iconic peptides for type 7 hexon protein, select 10 iconic peptides for type 55 hexon protein (peptide length is 8-15 amino acid residues, COOH terminal and NH 2 The end is lysine or arginine residues); by artificially synthesizing the full-length sequences of the above-mentioned various types of hexon genes of adenoviruses and corresponding landmark peptides in series to add lysine or arginine genes to the gene junctions, by PCR was combined with molecular cloning technology to import and export the target fragment into Escherichia coli BL21 (DE...

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Abstract

The invention relates to a method for detecting type-5, type-7 and type-55 adenoviruses by using a mass spectrum multiple reaction monitoring technology. The method comprises the following specific steps: synthesizing specific peptide fragment genes of hexon proteins of type-5, type-7 and type-55 adenoviruses; preparing a labeled peptide fragment mixed reagent; determining an ion peak value of the diagnostic marker; preparing the kit; and detecting the sample. The invention provides a new marker for early warning and diagnosis of adenovirus type, type 7 and type 55 infection, and has important clinical value. A mass spectrum result is utilized to confirm that mass spectrum ion peaks consistent with the standard and the heavy standard of preset adenovirus type 5, type 7 and type 55 hexon protein symbolic peptide fragments appear in a serum sample of a patient in the early stage of adenovirus infection. The SILAC-labeled winning and heavy-labeled hexon protein standard mixed reagent of adenovirus type 5, type 7 and type 55 can be used as an early warning and diagnosis marker for adenovirus type, type 7 and type 55 infection, and the result is accurate.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a method for detecting type 5, 7 and 55 adenoviruses by using mass spectrometry multiple reaction monitoring technology. Background technique [0002] Adenovirus (adenovirus) is a non-enveloped particle with a diameter of 70-90 nm, which is composed of 252 particles arranged in an icosahedron. Among them, the hexon (hexon, 120kD) composed of 240 non-vertex capsomers is the main antigenic protein of the adenovirus capsid, and each hexon is composed of 3 subunits, containing a large number of specific antigenic epitopes and Neutralizing epitopes, which can stimulate the body to produce type-specific neutralizing antibodies, are also criteria for diagnosing different serotypes. The specific serotype is closely related to the severity and clinical manifestations of the disease, so the detection and typing of adenovirus is more important. At present, there are 67 different serotypes of adeno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848G01N2333/075
Inventor 徐忠伟朱文清江涛杨震赵爱源张妍邹爽王天添
Owner LOGISTICS UNIV OF CAPF
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