Colloidal gold immunochromatography test paper for detecting cat toxoplasma gondii infection and preparation method thereof

An immunochromatographic test strip and Toxoplasma gondii technology, which is applied in the field of immunology, can solve the problems of inappropriate use of grassroots on-site operation and long detection time, and achieve the effects of fast detection speed, low cost, and no need for instruments and equipment.

Inactive Publication Date: 2017-12-12
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The test requires special test personnel, related instruments and equipment, long test time and other reasons, so it is not suitable for on-site operation at the grassroots level

Method used

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  • Colloidal gold immunochromatography test paper for detecting cat toxoplasma gondii infection and preparation method thereof
  • Colloidal gold immunochromatography test paper for detecting cat toxoplasma gondii infection and preparation method thereof
  • Colloidal gold immunochromatography test paper for detecting cat toxoplasma gondii infection and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. Expression and purification of surface antigen SAG3 recombinant protein

[0038] 1. Induced expression of recombinant protein SAG3

[0039] (1) Take 3 μL of prokaryotic-expressed SAG3 bacterial liquid, 5 μL of kanamycin in 5 mL of LB medium, transfer to a 37°C constant temperature shaking incubator, 150r / min, and shake overnight;

[0040] (2) Transfer the bacterial liquid shaken overnight to 300 mL of LB medium, add 300 μL of kanamycin at the same time, seal and transfer to a 37°C constant temperature shaking incubator for constant temperature shaking culture, 1.5h~2h;

[0041] (3) Add 300 μL of IPTG for induction, centrifuge at 7000r×7min at 4°C after 4~5h, and collect the bacterial liquid;

[0042] (4) Take PBS to resuspend the bacterial solution collected in step 3), ultrasonically break, and centrifuge to discard the supernatant;

[0043] (5) Dissolve the precipitate with 2mol / L urea and centrifuge, discard the supernatant; repeat more than 5 times;

[0044] (...

Embodiment 2

[0054] Preparation of Colloidal Gold Test Strips for Immunity of Cat Toxoplasma gondii Infection

[0055] (1) Preparation of colloidal gold solution

[0056] Choose trisodium citrate reduction method for preparation, take 99mL tetradistilled water, add 1mL chloroauric acid solution (1%), add 1mL trisodium citrate solution (1%), and prepare 40nm colloidal gold particles.

[0057] (2) Identification of colloidal gold particles

[0058] The colloidal gold was identified by transmission electron microscopy, and the colloidal gold solution was dropped on a nickel grid, dried with filter paper, and then observed by an electron microscope. High-quality colloidal gold particles are uniform in size, distributed in dispersed form without aggregation.

[0059] (2) Determination of the optimal pH of the gold-labeled antibody

[0060] Using 0.2mol / L K 2 CO 3 solution to adjust the pH of the colloidal gold solution, and judge by observing the color change.

[0061] (3) Preparation of ...

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PUM

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Abstract

The invention discloses a colloidal gold immunochromatographic test paper for detecting toxoplasma gondii infection in cats, and provides the same and a preparation method for serum detection of toxoplasma gondii infection in cats. The prepared colloidal gold test paper for toxoplasma gondii infection in cats has simple, Fast, no special equipment and other characteristics. The test results are easy to interpret and are suitable for rapid clinical diagnosis and large-scale epidemiological investigation. The indirect colloidal gold detection method is used, wherein the purified recombinant protein SAG3 ensures the specificity of the present invention. Use Staphylococcus aureus (SPA) for labeling, which is readily available and inexpensive. Compared with the traditional indirect hemagglutination test and indirect immunofluorescence test, the detection speed is faster, avoiding contact with live insects, and ensuring the safety of testing personnel.

Description

technical field [0001] The invention discloses a colloidal gold immunochromatographic test paper for detecting toxoplasma gondii infection in cats, and provides the same and a preparation method for serum detection of toxoplasma gondii infection in cats, belonging to the technical field of immunology. Background technique [0002] Toxoplasmosis is an important zoonotic parasitic disease caused by Toxoplasma gondii. Cats are the ultimate host of the disease and also an intermediate host. Cats infected with Toxoplasma gondii can infect humans and other intermediate hosts such as animals by excreting feces with infectious oocysts. OIE lists toxoplasmosis as a category B zoonotic disease. IHA and IFA are the detection methods stipulated in the national standard. However, the "national standard" method has many disadvantages such as complicated operation, need of professionals, time-consuming, high detection conditions, etc., and is not suitable for widespread promotion in pra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558
CPCG01N33/558G01N2333/45
Inventor 宫鹏涛刘维建杨正涛张西臣李建华杨举李赫李棕松
Owner JILIN UNIV
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