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Recombinant protein of human echinococcosis antigen and application thereof

A recombinant protein and hydatid disease technology, applied in the field of immunomedicine, can solve problems such as difficulty in protein selection, achieve high specificity and sensitivity, facilitate large-scale screening of human echinococcosis, and improve specificity and sensitivity.

Pending Publication Date: 2022-02-22
BGI GENOMICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If there are very few types of echinococcosis proteins identified, it will be more difficult to select proteins that may be used as echinococcosis antigens

Method used

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  • Recombinant protein of human echinococcosis antigen and application thereof
  • Recombinant protein of human echinococcosis antigen and application thereof
  • Recombinant protein of human echinococcosis antigen and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Identification of hydatid protein from hydatid cysts isolated after operation of hydatid patients

[0036] 1) Firstly, the hydatid cysts isolated from 6 hydatid patients (the first batch) were divided into four parts according to their tissue structure to extract protein. From the inside to the outside, they are protoscoleum, cyst fluid, germinal layer and cuticle. ; According to whether the cyst fluid is transparent or not, the 6 hydatid cysts were divided into a transparent cyst fluid group and a non-transparent cyst fluid group.

[0037] 2) Protein extraction was performed on the four components of each hydatid cyst, and the extracted protein was subjected to liquid enzymatic hydrolysis, followed by LC-MS / MS identification using a QE mass spectrometer;

[0038] 3) The original data of the QE off-machine use maxquant protein identification software, use human and hydatid protein database collections, carry out hydatid protein identification, statistical identificat...

Embodiment 2

[0083] This embodiment uses the MG2 recombinant protein as described in Example 1, and the detection objects are 60 cases of B-ultrasound positive plasma (including suspected) samples and 33 cases of normal human plasma (determined as negative) samples, according to the standard operating procedures of indirect ELISA Implement validation.

[0084] 1. ELISA experimental operation steps

[0085] 1) Antigen quantification: Each antigen was blown evenly with a pipette gun before coating, and a micro-ultraviolet spectrophotometer was used to quantify the antigen to ensure that the protein was not degraded. If there is no absorption peak at A280, it means that the amount of antigen is very low and it is not suitable for coating; if the protein is precipitated or insoluble, add 8M urea dropwise and blow evenly, centrifuge to get the supernatant and use the Bradford method to re-quantify the protein concentration.

[0086] 2) Coating: compare the concentration of the antigen tube wal...

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Abstract

The invention discloses a recombinant protein of a human echinococcosis antigen and an application of the recombinant protein. The recombinant protein comprises one or more of amino acid sequences as shown in SEQ ID NO: 1-10. The mutated amino acid sequence has at least 80%, 85%, 90%, 95%, 98% and 99% identity with the amino acid sequence of the recombinant protein, and maintains the function of the recombinant protein. The immune response rate of the recombinant protein of the human echinococcosis antigen to echinococcosis patients reaches 100%, and the recombinant protein can be used for manufacturing an ELISA kit and clinically detecting human echinococcosis. The single use or combined use of the recombinant protein may improve the specificity and sensitivity of laboratory diagnosis of echinococcosis.

Description

technical field [0001] The invention belongs to the field of immunomedicine, and in particular relates to a recombinant protein of human echinococcosis antigen and its application. Background technique [0002] Hydatid disease, also known as echinococcosis, is a zoonotic disease, especially in central and western China (Tibet, Qinghai, Sichuan, Xinjiang, etc.) infection. Human echinococcosis is mainly divided into cystic echinococcosis and alveolar echinococcosis. Imaging detection is the most intuitive and commonly used diagnostic method for echinococcosis. However, due to the long incubation period of echinococcosis, and the cysts formed by echinococcosis can only be detected by imaging when the cysts formed by echinococcosis are large enough, serum Medical detection is one of the important auxiliary means for imaging diagnosis of echinococcosis. At present, the serological diagnostic antigens that can be used to diagnose echinococcosis mainly include Antigen B (Antigen ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/70C12N1/21G01N33/68A61K39/00A61P33/10C07K16/18C12R1/19
CPCC07K14/43554C12N15/70G01N33/6893A61K39/0003A61P33/10C07K16/18G01N2333/43543A61K2039/53Y02A50/30
Inventor 张聪敏任艳刘斯奇
Owner BGI GENOMICS CO LTD
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