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Recombinant antigen for detecting schistosoma japonicum katsurada as well as preparation method and application of recombinant antigen

A technology for recombinant antigens and schistosomes, applied in the biological field, can solve problems such as low sensitivity, insufficient sensitivity, and long detection time

Pending Publication Date: 2021-06-29
长春万成生物电子工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing indirect hemagglutination assay (IHA) (ELISA) still generally has the problems of long detection time and insufficient sensitivity, while the antigens used in detection methods based on immunoenzyme-linked adsorption assay and immunocolloidal gold technology are generally GST, which is less immunogenic and less sensitive

Method used

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  • Recombinant antigen for detecting schistosoma japonicum katsurada as well as preparation method and application of recombinant antigen
  • Recombinant antigen for detecting schistosoma japonicum katsurada as well as preparation method and application of recombinant antigen

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Experimental program
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Effect test

Embodiment 1

[0027] Recombinant antigen 1 comprises GST protein fragment (SEQ ID NO.1), rSjGCP protein fragment (SEQ ID NO.2) and Sj23 (SEQ ID NO.3) protein fragment of Schistosoma japonicum connected sequentially from N-terminal to C-terminal, said The GST protein fragment and the rSjGCP protein fragment are connected through the connecting peptide 1 (SEQ ID NO.4: SDLEVLFQGPRGS) with the amino acid sequence shown in SEQ ID NO.4, and the rSjGCP protein fragment and the Sj23 protein fragment are connected through the amino acid sequence shown in SEQ ID NO. The connecting peptide 2 (SEQ ID NO.5: GGGGSGGGGS) shown in .5 is connected, and its amino acid sequence is shown in SEQ ID NO: 6:

[0028] MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLEVLFQGPRGSMRIGYEGLPRDQWPKVIHWNLHARDGIIWVLDGLLKCPEKLCPLLAEDVDYYGGGGSGGGGSMTGALENPNE...

Embodiment 2

[0036] 50 rabbits were taken, infected with 300 Schistosoma japonicum, and then treated with praziquantel (80 mg / kg body weight) 6 weeks after infection. Blood samples were collected before infection, 2 weeks, 4 weeks, and 6 weeks after infection, and 2 weeks, 4 weeks, and 6 weeks after praziquantel treatment. In addition, the serum of 20 rabbits infected with Fasciola gigantea was collected by the same method as a reference substance, and the serum of 20 rabbits not infected was collected as a negative control substance. Add 100 ul of PBS (PH=7.6) containing recombinant antigen 1 shown in SEQ ID NO.7, recombinant antigen 2 or GST protein shown in SEQ ID NO.8 at a concentration of 20 ug / ml in a polystyrene 96 microwell plate Coating was performed overnight at 4°C, and then rabbit serum diluted 1:100 was added to each well and incubated at 37°C for 1 hour. Add goat anti-mouse IgG-coupled horseradish peroxidase antibody and incubate at 37°C for 3 hours, then add 200ul of PBS co...

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Abstract

The invention relates to the technical field of biology, in particular to a recombinant antigen for detecting schistosoma japonicum katsurada. The recombinant antigen comprises a GST protein fragment, an rSjGCP protein fragment and an Sj23 protein fragment of the schistosoma japonicum katsurada, which are sequentially connected from an N end to a C end. Compared with the existing GST antigen, the recombinant antigen disclosed by the invention has the advantages that the immunogenicity of the antigen is improved to the maximum extent, the detection sensitivity is improved, the water solubility of the recombinant antigen is relatively high, and the purification efficiency is relatively high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant antigen for detecting Schistosoma japonicum, its preparation method and application. Background technique [0002] Schistosoma japonicum, also known as Schistosoma japonicum, has three main types of schistosomes that parasitize humans: Schistosoma haematobium, which is prevalent in northern Africa; Schistosoma mansoni, which is prevalent in Latin America and central Africa, and Schistosoma japonicum, which is prevalent in Asia. damage. [0003] At present, the methods currently used for the diagnosis of schistosomiasis mainly include etiological diagnosis, immunological diagnosis and ultrasonic diagnosis. The traditional etiological diagnosis method is the most reliable and classic diagnosis of schistosomiasis, and it is the gold standard for judging the prevalence of schistosomiasis, especially the Kato-Katz smear feces test and the miracidium incubation method MHT, b...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N9/10C12N15/62C12N15/70C12N1/21G01N33/68C12R1/19
CPCC07K14/43559C12N9/1088C12N15/70G01N33/6854C12Y205/01018C07K2319/40C07K2319/61G01N2333/43547
Inventor 王竑婷王立坚王晓明
Owner 长春万成生物电子工程有限公司
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