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Porcine trichinosis antibody test strip, and preparation method and application thereof

An antibody detection, Trichinella suis technology, applied in the measurement device, through the chemical reaction of the material for analysis, fluorescence/phosphorescence and other directions, can solve the problems of complex ES antigen components, cross-reaction in the diagnostic blind area, hindering practical application, etc. Shorten the detection blind area, has the effect of market development value and strong practicability

Inactive Publication Date: 2019-09-10
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ES antigen has complex components, cumbersome preparation, long production cycle, uneven batch quality, and serious diagnostic blind spots (cannot be detected before 19 days after infection) and cross-reactivity, which hinder its practical application.

Method used

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  • Porcine trichinosis antibody test strip, and preparation method and application thereof
  • Porcine trichinosis antibody test strip, and preparation method and application thereof
  • Porcine trichinosis antibody test strip, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1. Pig trichinellosis antibody detection test strip.

[0031]The pig trichinellosis antibody detection test strip described in this embodiment includes a sample pad, a binding pad, a chromatographic membrane, a water-absorbing pad and a bottom plate, and the binding pad is marked with time-resolved fluorescent microspheres coupled with goat anti-pig IgG; The sample pad is provided with a sampling hole; the chromatographic membrane is provided with a detection line and a quality control line, wherein the detection line is sprayed with a Trichinella spiralis antigen, and the Trichinella spiralis antigen is composed of Ts-WM5 in the muscle larval stage. Antigen, Ts-WN10 antigen of enteric infectious larvae, Ts-ZH68 antigen of adult stage and Ts-T668-C antigen of newborn larvae stage (T688 C-terminal immunodominant region) composed of four recombinant antigens. The Ts-WM5 recombinant antigen in the prokaryotic expression of the Trichinella spiralis cocktail antig...

Embodiment 2

[0032] Example 2. Preparation method of swine trichinellosis antibody detection test strip.

[0033] 1. Preparation of Trichinella spiralis cocktail antigen: Ts-WM5 recombinant protein expressed in muscular larvae, Ts-WN10 recombinant protein in intestinal infectious larvae, Ts-ZH68 recombinant protein in adult stage and Ts-T668 in neonatal larvae stage Each recombinant plasmid of the -C recombinant protein was transformed into host bacteria, and after induced expression, the bacterial cells were disrupted by ultrasonication, the precipitate was collected by centrifugation, and the inclusion bodies were dissolved with urea, and then the Ts-WM5 recombinant antigen in the muscle larval stage and the intestinal tract were respectively obtained by affinity purification. Ts-WN10 recombinant antigen of infectious larvae, Ts-ZH68 recombinant antigen of adult stage, Ts-T668-C recombinant antigen of neonatal larval stage, each recombinant antigen was mixed according to concentration rat...

Embodiment 3

[0119] Embodiment 3. The preparation method of swine trichinosis detection test strip.

[0120] Repeat Example 2, the difference from Example 2 is that the Trichinella spiralis cocktail antigen used as the detection line reagent in step 3 in this implementation, wherein the Ts-WM5 antigen of the muscle larval stage, the Ts-WN10 antigen of the intestinal infectious larvae, Ts-ZH68 antigen in the adult stage and Ts-T668-C antigen in the neonatal larval stage, the final concentrations were 0.5mg / mL, 0.5mg / mL, 0.75mg / mL and 0.75mg / mL; rabbits used as quality control line reagents Anti-goat IgG at a concentration of 0.75 mg / mL. The detection method described in Example 2 was used to investigate the sensitivity, specificity and stability of the swine trichinellosis antibody detection test strip prepared in this embodiment. The results showed that detection at this concentration was the same as ES ELSIA. Can improve detection sensitivity, reach early stage and detect trichinella spi...

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Abstract

The present invention relates to a porcine trichinosis antibody test strip, a preparation method and application thereof, and belongs to the technical field of fluorescence immunoassay. In order to detect trichinella infection more quickly, stably and accurately, the present invention provides a porcine trichinosis antibody test strip. The porcine trichinosis antibody test strip comprises a samplepad, a binding pad, a chromatographic membrane, an absorbent pad and a bottom plate; the binding pad is marked with time-resolved fluorescent microspheres coupled with the goat anti-porcine IgG; andthe chromatographic membrane is provided with a detection line and a quality control line, wherein the detection line is sprayed with the trichinella cocktail antigen composed of a recombinant musclelarval stage Ts-WM5 antigen expressed by prokaryotic cells, an intestinal infectious larval stage Ts-WN10 antigen, an adult stage Ts-ZH68 antigen and a newborn larval stage Ts-T668-C antigen, and thequality control line is sprayed with the rabbit anti-goat IgG. The test strip is prepared after combination of the components. The technical scheme of the present invention can be used for early rapiddetection of porcine trichinella infection.

Description

technical field [0001] The invention belongs to the technical field of fluorescent immunodetection, and in particular relates to a swine trichinellosis antibody detection test strip and a preparation method and application thereof. Background technique [0002] Trichinellosis is a very serious zoonotic disease. It not only causes huge economic losses to animal husbandry production, but also poses a huge threat to human health. People or animals mainly contain trichinosis through raw or semi-raw food. Caterpillar meat (mainly pork) and disease. [0003] For the inspection of trichinellosis in slaughtered animals, the commonly used legal inspection methods are microscope inspection and sample digestion method. However, the above two methods have certain disadvantages. Microscopic examination is time-consuming and laborious and has poor sensitivity. The sensitivity can only be detected when the density of worms in the meat reaches 3 worms per gram. Although the collection and...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558G01N33/543G01N33/533G01N21/64G01N21/78
CPCG01N21/6408G01N21/78G01N33/533G01N33/54313G01N33/558G01N33/569G01N2021/6439
Inventor 刘明远刘晓雷杨勇王学林白雪唐斌丁静王楠张小波
Owner JILIN UNIV
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