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An immunocolloidal gold test paper strip for detecting a pig toxoplasmosis circulating antigen and a preparing method thereof

A technology of colloidal gold test paper and toxoplasmosis, which is applied in the field of serological detection, achieves the effect of strong specificity and simple operation

Inactive Publication Date: 2017-11-17
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it has not been reported that the monoclonal antibody coated with NC membrane T-line assembly test strips prepared by using the surface antigen SAG3 of Toxoplasma gondii and used to detect the serum circulating antigen of Toxoplasma gondii

Method used

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  • An immunocolloidal gold test paper strip for detecting a pig toxoplasmosis circulating antigen and a preparing method thereof
  • An immunocolloidal gold test paper strip for detecting a pig toxoplasmosis circulating antigen and a preparing method thereof
  • An immunocolloidal gold test paper strip for detecting a pig toxoplasmosis circulating antigen and a preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Preparation of colloidal gold probes

[0054] The specific preparation method of the immune colloidal gold detection test strip for the detection of swine toxoplasma gondii serum circulating antigen is as follows:

[0055] (1) Preparation of colloidal gold

[0056]Perform the following treatments on the glassware used: fully rinse, soak in acid, rinse again, and dry for later use. A colloidal gold solution with a particle size of 40 nm was prepared by using a trisodium citrate reduction method. Put the rotor in the flask, add 99mL of deionized water, weigh it with an electronic balance and record it, heat it with a constant temperature magnetic stirring electric heating mantle, turn on the rotor switch when there are large bubbles on the rotor, and start adding 1mL of 1% chlorine gold acid solution. Add 1% trisodium citrate solution when large bubbles appear again, observe the color change of the solution in the flask at this time, and start timing for 5-10 minutes w...

Embodiment 2

[0065] Assembling and testing test strips

[0066] (1) Handling of the sample pad

[0067] Soak the sample pad in the sample pad treatment solution (containing 1% BSA, 0.5% S17 and 0.5% tx10) for 10 minutes, take it out and dry it in a 37°C incubator, dry it at 4°C and store it sealed.

[0068] (2) Preparation of gold standard pad

[0069] Cut the gold label pad into a size of 0.8cm×3.2cm. The identified high-quality gold-labeled antibody was added dropwise and spread evenly on the cut gold-labeled pad, placed in a 37°C incubator to dry for 2-3 hours, and then dried and sealed at 4°C.

[0070] (3) Coated T-line and C-line

[0071] Add antibody to the NC membrane of the assembled test strip to coat the T and C lines, add 1mg / mLAnti-A-SAG3-23 1μl at a distance of 5-6mm from the gold standard pad, this is the T line, and then distance Add 1 μl of 1mg / mL goat anti-mouse IgG antibody dropwise at the 8-10mm of the T line, which is the C line, and store it at 4°C after being full...

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Abstract

An immunocolloidal gold test paper strip for detecting a pig toxoplasmosis circulating antigen and a preparing method thereof are provided. The colloidal gold immunochromatography technique is utilized to detect the circulating antigen in blood of pigs affected with toxoplasmosis. T line coating and gold marking pad preparation utilize two monoclonal antibodies resisting a toxoplasma gondii surface antigen SAG3 respectively. The test paper strip has no cross reaction with two types of pig cysticercus positive serum so that the test paper strip has high specificity. The test paper strip has biosecurity. Preparation of the gold marking pad and coating of a T line apply the two monoclonal antibodies resisting the toxoplasma gondii surface antigen SAG3, a goat anti-mouse antibody is coated by a C line, the antibodies are nontoxic, and the test paper strip does not cause environment pollution.

Description

technical field [0001] The invention provides an immune colloidal gold test strip for detecting swine toxoplasmosis circulating antigens and a preparation method thereof. The colloidal gold immunochromatography technique is used to detect circulating antigens in swine toxoplasma gondii blood, and belongs to the technical field of serological detection. Background technique [0002] Toxoplasmosis ( Toxoplasmosis ) is produced by Toxoplasma gondii ( Toxoplasma gondii ) caused by a serious zoonotic parasitic disease, toxoplasmosis has brought serious impact on human health and the development of pig industry. Humans are infected with toxoplasmosis mainly by eating undercooked meat containing toxoplasma cysts. Therefore, the diagnosis of toxoplasmosis in pigs has public health significance. Among the existing diagnostic methods for toxoplasmosis, the etiological diagnosis results are indeed reliable, but if the parasites are not typical, misdiagnosis and missed diagnosis wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/569G01N33/558
CPCG01N33/558G01N33/56905G01N33/68
Inventor 张西臣杨正涛寇金华宫鹏涛李建华杨举李赫李棕松高珺珊
Owner JILIN UNIV
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