Immunogenic protein in schistosoma japonicum soluble egg antigen as well as screening method and application thereof

A technology of immunogenicity and screening method, applied to the field of immunogenic protein in Schistosoma japonicum soluble egg antigen and its screening

Inactive Publication Date: 2011-06-01
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the immunomics research on the soluble egg antigen of Schistosoma japonicum

Method used

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  • Immunogenic protein in schistosoma japonicum soluble egg antigen as well as screening method and application thereof
  • Immunogenic protein in schistosoma japonicum soluble egg antigen as well as screening method and application thereof
  • Immunogenic protein in schistosoma japonicum soluble egg antigen as well as screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Screening of immunogenic protein in soluble egg antigen of Schistosoma japonicum

[0041] 1. The detailed steps of two-dimensional electrophoresis combined with West blot are as follows:

[0042] 1. Collect sera from patients with schistosomiasis before and after praziquantel treatment. Miaoling Village, Daguan District, Anqing City, Anqing City, Anhui Province, an endemic area of ​​schistosomiasis, was used as an on-site investigation point to collect serum samples needed for the experiment. The resident residents of the village were randomly selected as the research objects, and fresh fecal samples were collected twice in a row for 2 consecutive days. According to the Kato-Katz fecal smear method [Katz N, Chaves A, Pellegrino JA simple device for quantitative stool thick-smear technique in Schistosomiasismansoni.1972, Rev Inst Med Trop Sao Paulo 14: 397-400] To make 6 smears with a fecal volume of 41.7 mg (second delivery and six inspection), cover and press the...

Embodiment 2

[0128] Example 2 Use of immunoenzyme-linked adsorption test to verify the selected diagnostic targets for schistosomiasis japonica

[0129] 1. Detailed steps of the experiment

[0130] 1. Select Schistosoma japonicum Heat Shock Protein 60 (Sj HSP60) from the immunogenic proteins of the soluble egg antigen of Schistosoma japonicum for prokaryotic expression and purification (refer to Merck pET Prokaryotic Expression Manual for specific operation methods).

[0131] 2. The purified sufficient protein is respectively 0.625μg / ml, 0.125μg / ml, 0.25μg / ml, 0.5μg / ml, 1.0μg / ml, 5.0μg / ml, 10μg / ml, 20μg / ml and 30μg / The concentration of ml was coated with 96-well microtiter plate at 4°C overnight.

[0132] 3. Wash the plate with PBST on a plate washer for 3 times, 3 minutes each time; add 100 μl of 1% BSA (bovine serum albumin) to each well, and block at 37°C for 2 hours.

[0133] 4. Wash the plate on the plate washer with PBST 3 times, 3 minutes each time; add 100μl / well of mixed patient serum (mi...

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Abstract

The invention discloses an immunogenic protein in a schistosoma japonicum soluble egg antigen. The immunogenic protein is selected from amino acid sequences shown in SEQ ID NO:2-SEQ ID NO:14. In addition, the invention also discloses a screening method and application of the immunogenic protein in the schistosoma japonicum soluble egg antigen and is mainly characterized by screening the immunogenic protein in the schistosoma japonicum soluble egg antigen through a combined proteomics technology and a serological method, particularly analyzing the immunogenic protein in the schistosoma japonicum soluble egg antigen by utilizing bidirectional electrophoresis, Westernblot (2D-WB) and a mass spectrum method and using the immunogenic protein as a potential target spot for schistosomiasis diagnosis.

Description

Technical field [0001] The present invention relates to the field of biological detection, in particular to combining protein science technology and serological technology to screen the immunogenic protein in the soluble egg antigen of Schistosoma japonicum and use it as a potential target for diagnosis of schistosomiasis japonicum, in particular to a soluble Schistosoma japonicum The immunogenic protein in the egg antigen and its screening method and application. Background technique [0002] As a global public health problem, schistosomiasis is the second largest parasitic disease in the world after malaria. Schistosomiasis has a long history of widespread distribution, and has seriously affected human health and social and economic development. It is estimated that schistosomiasis is endemic in 76 countries and regions in the world, with 200 million infected people, 500-600 million threatened people, of which about 120 million are symptomatic, about 20 million are severely il...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435G01N33/561G01N33/53
Inventor 胡薇鞠川徐斌卢艳张颋莫筱瑾陈绅波魏刚刚冯正
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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