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Sarcocystis fusion antigen, coding gene, indirect ELISA antibody detection kit and application thereof

A Sarcocystis, fusion antigen technology, applied in the field of animal or human disease detection, can solve the problems of cloning, expression and application development that have not yet been reported, and achieve good application prospects, strong antigenicity and specificity.

Active Publication Date: 2020-03-27
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no research reports on the cloning, expression and application development of the surface antigen SAG3 / 4 gene sequences of S. micheni and S. cruzi.

Method used

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  • Sarcocystis fusion antigen, coding gene, indirect ELISA antibody detection kit and application thereof
  • Sarcocystis fusion antigen, coding gene, indirect ELISA antibody detection kit and application thereof
  • Sarcocystis fusion antigen, coding gene, indirect ELISA antibody detection kit and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040] In the present embodiment, the fusion antigen of Sarcocystis is composed of the following three parts:

[0041] 1) Peptide 1 derived from the surface antigen SAG3 of Sarcocystis militaris;

[0042] 2) Peptide 2 derived from the surface antigen SAG4 of S. cruzi;

[0043] 3) A flexible peptide segment used to connect peptide segment 1 and peptide segment 2;

[0044] The amino acid sequence of the surface antigen SAG3 of S. mieni is shown in SEQ ID NO.10;

[0045] The amino acid sequence of the surface antigen SAG4 of S. cruzi is shown in SEQ ID NO.11;

[0046] The amino acid sequence of the flexible peptide is: GGGGSGGGGSGGGGS. The flexible peptide prevents the two connected protein peptides from interacting with each other when they are folded to form a high-level structure, maintaining strong antigenicity.

Embodiment 2

[0048] The amino acid sequence of the sarcocystis fusion antigen in the present embodiment is:

[0049] (as shown in SEQID NO.4).

[0050] In this sequence, the part in italics is the partial amino acid sequence of S. cruzi SAG3 (peptide segment 37-118 in the sequence SEQ ID NO.10), the underlined part is the flexible peptide, and the bold part is S. cruzi SAG4 Partial amino acid sequence (peptide segment 165-264 in sequence SEQ ID NO.11).

Embodiment 3

[0052] The nucleotide sequence of the gene encoding the sarcocystis fusion antigen (shown in SEQ ID NO.4) in this example is shown in SEQ ID NO.5. The sequence is obtained by referring to the codon usage frequency table of Escherichia coli and using codon optimization tools. The coding region of the fusion antigen is 597 bp, and the coding region of the bivalent fusion antigen SAG3-4 is artificially synthesized by a biological company.

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Abstract

The invention discloses a sarcocystis fusion antigen, a coding gene, an indirect ELISA antibody detection kit and application thereof, and belongs to the technical field of animal or human epidemic disease detection. The sarcocystis fusion antigen consists of a part of amino acid sequences (37 th-118 th sites) of a surface antigen SAG3 of sarcocystis miescheriana, 15 amino acid flexible peptides and a part of amino acid sequences (165 th-264 th sites) of a surface antigen SAG4 of sarcocystis cruzi. The fusion antigen has very high antigenicity and specificity, can generate effective antigen-antibody reaction with sarcocystis miescheriana and sarcocystis cruzi antibodies as a detection antigen, cannot generate cross immune reaction with positive serum of toxoplasma gondii and neospora caninum, is an ideal detection antigen for detecting the sarcocystis miescheriana and sarcocystis cruzi antibodies, and has good application prospects.

Description

technical field [0001] The invention relates to a sarcocystis fusion antigen, a gene encoding the fusion antigen, an indirect ELISA antibody detection kit containing the fusion antigen and an application thereof, belonging to the technical field of animal or human disease detection. Background technique [0002] Sarcocytosis is an important food-borne zoonotic parasitic disease. The asexual reproduction stage of Sarcocystis mainly parasitizes in the vascular endothelial cells of various organs of the intermediate host, the muscle fibers of striated muscle and cardiac muscle and the central nervous system, and the sexual reproduction stage mainly parasitizes in the small intestine of the final host. Animals such as pigs and cattle have a high infection rate, and Sarcocystis miescheriana and S. cruzi are the dominant and highly pathogenic species of pigs and cattle, respectively. Sick animals mainly show symptoms such as fever, anemia, emaciation, difficulty in movement and b...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N1/21G01N33/68C12R1/19
CPCC07K14/43504C07K2319/00C12N15/70G01N33/68
Inventor 闫文朝韩利方钱伟锋张旻位治国吕超超王天奇
Owner HENAN UNIV OF SCI & TECH
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