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Toxoplasma immune-gold labeled quick detection reagent and its preparation method

A technology for detecting reagents and immunogold standards, which is applied in the field of immunocolloidal gold reagents, can solve the problems of low-temperature storage of reagents, low sensitivity, complicated operation process, etc., and achieve high sensitivity and accuracy, convenient storage and transportation, and wide application range Effect

Inactive Publication Date: 2008-04-30
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of enzyme-linked immunosorbent assay are: special equipment such as microplate reader is required for use; reagents need to be stored at low temperature; detection operators need to be professionally trained; the operation process is relatively complicated, and the detection time is relatively long; The cost is high, and single person (head, only) testing cannot be realized; it is not conducive to the promotion and use in grassroots units
In addition to the above shortcomings, the indirect hemagglutination method also has the disadvantages of low sensitivity and prone to missed detection
Chinese invention patent applications CN1431513A and CN1431514A respectively disclose a "detection of toxoplasma antibody IgM immune colloidal gold reagent and preparation method" and a "detection of toxoplasma antibody IgG immune colloidal gold reagent and preparation method", they all have the following deficiencies (1) The preparation process of Toxoplasma-specific surface membrane antigen and monoclonal antibody is cumbersome; (2) The detection range of the reagent is limited, and it can only detect human toxoplasmosis; (3) The reagent cannot detect anti-Toxoplasma IgG simultaneously and IgM

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Preparation of Toxoplasma gondii metabolism and secretion antigen: On March 3, 2006, 20 mice with a body weight of 20-25 grams were inoculated intraperitoneally with 1×10^6 Toxoplasma gondii tachyzoites / only, and on March 5, the mice appeared After experiencing symptoms such as depression and rough coat, the mice were killed by taking off their necks, and then soaked in 75% alcohol to disinfect the body surface. Each mouse was washed with 2 ml of PH7.2 0.15mol / L PBS buffer solution to collect worms, and a total of 45 ml of peritoneal fluid was collected. The collected peritoneal fluid was centrifuged at 3000 rpm for 40 minutes, and the supernatant was collected. , put the supernatant in a refrigerator at -20°C, take out the supernatant from the refrigerator on March 7, centrifuge at a speed of 4,000 rpm for 3 times after melting, and centrifuge for 30 minutes each time, collect the supernatant, and detect the supernatant The protein content is 21.37 mg / ml, and the s...

Embodiment 2

[0025] (1) Preparation of Toxoplasma metabolic secretion antigen: On April 13, 2006, 20 mice with a body weight of 20-25 grams were inoculated intraperitoneally with 1×10^5 Toxoplasma gondii tachyzoites / only, and on April 17, the mice appeared After experiencing symptoms such as depression and rough coat, the mice were killed by taking off their necks, and then soaked in 70% alcohol to disinfect the body surface. Each mouse was washed with 2-3 ml of PH7.2 PBS buffer to collect worms. A total of 45ml of peritoneal fluid was collected, and the collected peritoneal fluid was centrifuged at a speed of 4,000 rpm for 30 minutes to collect the supernatant, which was placed in a refrigerator at -20°C. On April 18, the supernatant was taken out from the refrigerator and melted again Centrifuge twice at a speed of 3000 revolutions per minute, each time for 40 minutes, collect the supernatant, and detect that the protein content of the supernatant is 18.26 mg / ml. The supernatant is the ...

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Abstract

The invention relates to immunity gold mark quick testing reagent which measures the toxoplasma antibody and the preparative method thereof. One end of a PVC back lining is orderly stuck with a sample mat, a combination mat, a phroxylin film, and a sopping mat. The combination mat is wrapped with antigenic colloid gold combos metabolized and excreted by the toxoplasma, the top of the phroxylin film is wrapped with antigen metabolized and excreted by toxoplasma and toxoplasma antibodies. Through toxoplasma metabolizing antigen, preparing toxoplasma antibody, preparing colloid gold, preparing toxoplasma metabolizing and excreting antigen colloid gold mark craftworks, finally the toxoplasma metabolizing and excreting antigen colloid gold mark is wrapped on the colloid gold combination mat to wrap the toxoplasma metabolizing and excreting antigen and toxoplasma antibody in the measuring district and the sluggish control district in a line shape to produce the reagent. The invention with strong specificity, high sensitivity, simple operation, quick ability and cheap price is applicable to the filed measuring, the whole process only needs 20 minutes, the operators need not be professionally trained and the operation following the instruction can be fulfilled.

Description

technical field [0001] The invention relates to an immune colloidal gold reagent for detecting antibodies and a preparation method of the reagent. Background technique [0002] Existing methods for detecting antibodies to Toxoplasma gondii mainly include enzyme-linked immunosorbent assay and indirect hemagglutination method, both of which have defects and deficiencies. The disadvantages of enzyme-linked immunosorbent assay are: special equipment such as microplate reader is required for use; reagents need to be stored at low temperature; detection operators need to be professionally trained; the operation process is relatively complicated, and the detection time is relatively long; The cost is high, and single (head, only) detection cannot be realized; it is not conducive to popularization and use in grassroots units. In addition to the above shortcomings, the indirect hemagglutination method also has the disadvantages of low sensitivity and prone to missed detection. Chin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/544G01N33/532
Inventor 张德林王艳华李学瑞才学鹏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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