New echinococcosis antigen Murinoglobulin-2 protein

A protein and amino acid technology, applied in the fields of resistance to vector-borne diseases, peptide sources, instruments, etc., can solve problems such as insufficient specificity and/or sensitivity, difficulty in protein selection, and difficulty in isolating hydatid proteins

Active Publication Date: 2021-08-06
BGI GENOMICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, both natural purified antigens and recombinant purified antigens have certain deficiencies in specificity and / or sensitivity.
More importantly, the etiology of human echinococcosis involves factors such as difficulty in sampling and separation of echinococcosis protein, and there have been no more antigens that can be used for serological detection
Specifically, the prior art has the following deficiencies: 1) Extracting and identifying hydatid protein from hydatid cysts isolated after operation of hydatid patients is a technical difficulty in itself
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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  • New echinococcosis antigen Murinoglobulin-2 protein
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  • New echinococcosis antigen Murinoglobulin-2 protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1, the acquisition of echinococcosis neoantigen Murinoglobulin-2 protein

[0069] 1. The process of identifying more hydatid proteins from hydatid cysts isolated from hydatid patients after surgery

[0070] (1) Firstly, the hydatid cysts separated from 6 hydatid patients were divided into four parts according to their tissue structure to extract protein. Whether the cyst fluid is transparent or not, the 6 hydatid cysts were divided into a transparent group and an opaque group.

[0071] (2) The protein was extracted from the four components of each hydatid cyst, and the extracted protein was subjected to liquid enzymatic hydrolysis, and then identified by LC-MS / MS using a QE mass spectrometer.

[0072] (3) The raw data of QE off-machine was carried out using the maxquant protein identification software and the collection of human and hydatid protein databases (Unreviewed (TrEMBL) database downloaded from the Uniprot (https: / / www.uniprot.org / ) website) Hydatid p...

Embodiment 2

[0118] Example 2, ELISA Experimental Verification Case of Hydatid Recombinant Antigen Murinoglobulin-2 Recombinant Protein

[0119] The detected objects were 14 postoperative echinococcosis patient plasma (clinically confirmed as positive) and 6 cases of normal Tibetan plasma (clinically confirmed as negative), which were carried out according to the standard operating procedures of indirect ELISA, and the results showed that Murinoglobulin-2 recombinant protein (purified in Example 1, i.e. figure 2 The target protein eluted when the middle imidazole concentration is 250mM) the positive detection rate is 86%, and the negative detection rate is 100%; Minuo Biotechnology Co., Ltd., product number: YM-VI08) when testing the same plasma, the positive detection rate was 86%, and the negative detection rate was 100%.

[0120] 1. ELISA experiment specific operation steps

[0121] (1) Antigen quantification: Each antigen was blown evenly with a pipette gun before coating, and a mic...

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Abstract

The invention discloses a novel echinococcosis antigen Murinoglobulin-2 protein. The amino acid sequence of the Murinoglobulin-2 protein provided by the invention contains all or part of the following sites: the 31st site to the 39th site, the 89th site to the 96th site, the 107th site to the 115th site, the 179th site to the 200th site, the 203rd site to the 216th site, the 225th site to the 242nd site, the 255th site to the 273rd site, the 282nd site to the 300th site, the 309th site to the 315th site, the 336th site to the 354th site and the 379th site to the 390th site of SEQ ID No.1. The Murinoglobulin-2 protein provided by the invention can be used for clinically detecting the human echinococcosis. The Murinoglobulin-2 protein is used for detecting plasma of 14 cases of postoperative echinococcosis people and plasma of 6 cases of normal Tibetan people, the positive detection rate is 86%, and the negative detection rate is 100%. According to the invention, a hydatid antigen library is expanded, and a certain contribution is made to diagnosis of human-derived echinococcosis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the neoechinococcosis antigen Murinoglobulin-2 protein. Background technique [0002] Hydatid disease, also known as echinococcosis, is a zoonotic disease, and the infection is particularly serious in the central and western regions of China (Tibet, Qinghai, Sichuan, Xinjiang, etc.). 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. [0003] To date, several methods have been described for the laboratory diagnosis of echinococcosis, including the detect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/70C12N1/21G01N33/68
CPCC07K14/43554C12N15/70G01N33/6854G01N2333/43543Y02A50/30
Inventor 张聪敏任艳刘斯奇
Owner BGI GENOMICS CO LTD
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