Chemical luminescence immune analysis diagnosis reagent kit detecting Toxoplasma Gondi IgG antibody and preparation method thereof

A chemiluminescence immunoassay, diagnostic kit technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, analysis of materials, etc., can solve the problems of toxic substrates, inconvenient operation, low sensitivity, etc. High compliance, easy operation and good specificity

Inactive Publication Date: 2009-02-25
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] In order to solve the problems of inconvenient operation, low sensitivity, short validity period of radioimmunoassay reagents, pollution hazards of radioactive waste, interference of enzyme immunoassay reagents by various factors, and most toxic substrates in clinical etiology tests, the present invention provides a detection Chemiluminescent immunoassay diagnostic kit for Toxoplasma gondii IgG antibody

Method used

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  • Chemical luminescence immune analysis diagnosis reagent kit detecting Toxoplasma Gondi IgG antibody and preparation method thereof
  • Chemical luminescence immune analysis diagnosis reagent kit detecting Toxoplasma Gondi IgG antibody and preparation method thereof
  • Chemical luminescence immune analysis diagnosis reagent kit detecting Toxoplasma Gondi IgG antibody and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Preparation of a chemiluminescent immunoassay diagnostic kit for detecting IgG antibodies to Toxoplasma gondii:

[0069] A, coated plate preparation:

[0070] Coat the microwell plate with 20mmol / L pH7.4 phosphate buffer solution containing 2μg / mL FITC antibody at a volume of 100μL / well. Bovine serum, 5% (weight) sucrose 20mmol / L pH 7.4 phosphate buffer 120μL / well was sealed for 18-20 hours, dehumidified after shaking off the blocking solution, dried at 28°C for 18-20 hours, added a desiccant to seal and Check the sealed aluminum foil bag for air leakage and store at 2-8°C.

[0071] B, preparation of FITC-labeled Toxoplasma gondii antigen:

[0072] a), FITC labeling: dialyze 2 mg of Toxoplasma gondii antigen with 50 mmol / L pH9.6 carbonic acid buffer at 2-8°C, and dissolve FITC in dimethyl sulfoxide (DMSO) to a final concentration of 1 mg / mL , according to the ratio of Toxoplasma gondii antigen:FITC=1mg:150μg, slowly add FITC dimethyl sulfoxide solution into the Toxop...

Embodiment 2

[0096] Preparation of a chemiluminescent immunoassay diagnostic kit for detecting IgG antibodies to Toxoplasma gondii:

[0097] Using magnetic particles as a solid phase carrier, the FITC antibody was coupled to the surface of the magnetic particles by the EDC method, and concentrated with 12% (weight) NaCl, 0.1% (weight) KCl, 400mmol / L pH7.2 Tris-HCl buffer solution Washing solution and the rest were prepared in the same manner as in Example 1 to prepare the Toxoplasma gondii IgG antibody chemiluminescent immunoassay kit.

Embodiment 3

[0099] Preparation of a chemiluminescent immunoassay diagnostic kit for detecting IgG antibodies to Toxoplasma gondii:

[0100] (1), preparation of luminescent substrate solution:

[0101] a), chemiluminescent substrate solution A

[0102] The following formula was used: 1mmol / L luminol, 0.1mmol / L 4-hydroxybiphenyl, 0.01mmol / L 4-iodophenylboronic acid, 50mmol / L pH8.0~10.0 boric acid buffer. Add 0.177g of luminol, 0.017g of 4-hydroxybiphenyl, 0.0025g of 4-iodophenylboronic acid, 2.85g of boric acid, and 1.225g of borax into a clean container and dissolve them in double-distilled water. Adjust the pH value to 8.0-10. Distilled water to dissolve to 1000mL.

[0103] b), chemiluminescent substrate solution B

[0104] The following formula was adopted: 1mmol / L carbamide peroxide, 0.05% (volume) Tween 20, 10mmol / L pH7.0~7.6 phosphate buffer. 0.094g carbamide peroxide, 0.5ml Tween20, 25.79gNa 2 HPO 4 12H 2 O, 4.37g NaH 2 PO 4 2H 2 Add O into a clean container and dissolve wi...

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Abstract

The invention discloses a diagnostic kit for testing toxoplasma IgG antibody chemiluminescence immunoassay. The diagnostic kit comprises a negative reference substance, a positive reference substance, a solid phase carrier, a toxoplasma antigen marker, an enzyme combination, a chemiluminescence substrate solution A, a chemiluminescence substrate solution B and a concentrated washing solution. Moreover, the invention further discloses a preparation method for the diagnostic kit for testing toxoplasma IgG antibody chemiluminescence immunoassay. The kit has the characteristics of simple and convenient operation, high sensitivity, good specificity and extremely high conformity to etiological test result, etc. The invention can be taken as one of the important indexes for pregestational test, and is significant for improving the health of newborns and promoting good prenatal and postnatal care.

Description

technical field [0001] The invention relates to the field of medical immunological in vitro diagnostic reagents, in particular to a chemiluminescent immunoassay diagnostic kit for detecting IgG antibodies to Toxoplasma gondii and a preparation method thereof. Background technique [0002] Toxoplasma gondii is an intestinal coccidia of cats. It was discovered by French scholars Nicolle and Manceaux in the spleen mononuclear cells of Ctenodactylus gondii. Toxoplasma gondii. Toxoplasmosis, also known as toxoplasmosis, is a zoonotic parasitic disease caused by Toxoplasma gondii. According to literature reports, Toxoplasma gondii can infect a variety of mammals, and the infection rate of livestock can reach 10-50%. There are about 1 billion people infected with Toxoplasma gondii in the world, and the average infection rate of our population is 5.16%. [0003] Toxoplasma gondii is one of the five major pathogens causing fetal malformation caused by intrauterine infection of pre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/76
Inventor 彭京胜应希堂宋胜利胡国茂郑金来唐宝军于尚永
Owner CHEMCLIN DIAGNOSTICS CO LTD
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