Specific detection antigen for trichinosis and application of specific detection antigen

A technology for detecting lines and proteins, applied in applications, biological tests, measuring devices, etc., can solve the problems of difficult commercialization, expensive preparation, false positives, etc., and achieve the effects of simple operation, rapid diagnosis method, and good specificity.

Pending Publication Date: 2021-09-17
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These methods usually use excreted secretions as antigens, which are hig

Method used

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  • Specific detection antigen for trichinosis and application of specific detection antigen
  • Specific detection antigen for trichinosis and application of specific detection antigen
  • Specific detection antigen for trichinosis and application of specific detection antigen

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Embodiment 1, preparation of putative trypsin (Tsp_00436) recombinant antigen

[0037] This embodiment provides a preparation method for the preparation of putative trypsin (Tsp_00436) recombinant antigen. The amino acid sequence of putative trypsin (Tsp_00436) recombinant antigen optimized according to the putative trypsin (Tsp_00436) gene sequence is shown in SEQ ID NO.1, which encodes the protein The gene sequence of is shown in SEQ ID NO.2.

[0038]1. The gene fragment was synthesized by Suzhou Synbio Biotechnology Co., Ltd. with the expression vector putative trypsin (Tsp-00436) containing the corresponding target gene. The steps of double digestion are as follows:

[0039] (1) Centrifuge the centrifuge tube containing the lyophilized powder of the expression vector of the target gene at 3000 rpm / normal temperature for 1 min.

[0040] (2) with 50 μL sterile ddH 2 O Dissolve the lyophilized powder, then mix gently with a vortex instrument, and centrifuge for 30 se...

Embodiment 2

[0098] Embodiment 2, preparation of trichinellosis time-resolved fluorescent immunochromatography test strip

[0099] Such as image 3 As shown, the present embodiment provides a kind of trichinosis time-resolved fluorescence immunochromatography test paper, and this time-resolved fluorescence immunochromatography test paper comprises sample pad 2, binding pad 3, nitrocellulose film 5 (NC film, layer analysis film) and absorbent paper 4 and PVC bottom plate 1. Sample pad 2, binding pad 3, nitrocellulose membrane 5, absorbent paper 4 on PVC bottom plate 1. Wherein, one end of the nitrocellulose membrane 5 is laminated with one end of the binding pad 3, the other end of the nitrocellulose film 5 is laminated with one end of the absorbent paper 4, and one end of the sample pad 2 is laminated with the other end of the binding pad 3; 3 is coated with goat anti-bovine IgG antibody labeled with fluorescent microspheres, a detection line 6 is set on the side of the nitrocellulose fi...

Embodiment 3

[0149] Embodiment 3, time-resolved fluorescent microsphere chromatography test strip specificity

[0150] Test the positive serum samples of Trichinella spiralis sheep, Yangstomum sheep positive serum, Esophagostomum positive serum of sheep, and Sheeptail worm positive serum. Except for the positive serum of Trichinella spiralis sheep, other non-sheep No signal was detected on the T line of the serum of the Trichinella spiralis pathogen, and only a band appeared on the C line, which was negative, and the result was as follows: Figure 4 .

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Abstract

The invention discloses a specific detection antigen for trichinosis and application of the specific detection antigen. The antigen protein is a protein as shown in (a1) or (a2): (a1) a protein with an amino acid sequence as shown in SEQ ID No.1; and (a2) a protein which is obtained by substitution and/or deletion and/or addition of one or more amino acid residues on the amino acid sequence of the protein limited by (a1) and can be specifically combined with a sheep trichinosis serum antibody. According to the specific detection antigen for trichinosis and the application of the specific detection antigen, the detection time is advanced to 48 hours for the first time, the operation is simple and convenient, and the sensitivity and the accuracy are good; and the method established by the invention has good specificity, provides a rapid diagnosis method for epidemiological investigation and disease diagnosis, and can be used for simultaneously detecting serum in different stages of adult stage, newborn larva stage and muscle larva stage of infected trichina.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a specific detection antigen for trichinellosis and its application. Background technique [0002] Trichinellosis is a serious global zoonotic disease. Humans or ruminants are mainly infected by eating meat infected by Trichinella. The disease is a second-class animal disease, which causes huge economic losses to the animal husbandry industry loss. At present, regarding the diagnosis of Trichinella spiralis, the methods recognized by the International Veterinary Office (OIE) are tablet microscopy and digestion. However, the above two methods have certain limitations in the detection, and the microscopic examination is time-consuming and laborious and has low sensitivity. Although the digestion method can improve the detection rate of Trichinella spiralis to a certain extent, the detection rate of this method must be more than 3 worms per gram of meat before it can ...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12G01N33/68G01N33/558
CPCC07K14/4354G01N33/68G01N33/558G01N2333/4353
Inventor 杨娜桑晓宇王彦虎丁莹莹冯颖陈冉
Owner SHENYANG AGRI UNIV
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