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Reagent strip for testing syphilis specific IgM antibodies through gold immunochromatographic assay and preparation method thereof

A technology of immunochromatographic detection and antibody detection, which is applied in the direction of measuring devices, analytical materials, instruments, etc., to achieve the effects of strong specificity, high sensitivity, and low cost

Inactive Publication Date: 2010-10-13
ZHONGSHAN HOSPITAL XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a negative TP-IgM does not completely rule out infectivity, a positive TP-IgM must suggest that the patient is infectious

Method used

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  • Reagent strip for testing syphilis specific IgM antibodies through gold immunochromatographic assay and preparation method thereof
  • Reagent strip for testing syphilis specific IgM antibodies through gold immunochromatographic assay and preparation method thereof
  • Reagent strip for testing syphilis specific IgM antibodies through gold immunochromatographic assay and preparation method thereof

Examples

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preparation example Construction

[0034] The preparation method of the syphilis-specific IgM antibody colloidal gold immunochromatography detection reagent strip comprises the following steps:

[0035] 1) Preparation of recombinant syphilis antigens TPN17 and TPN47

[0036] Using gene cloning technology, PCR amplified DNA encoding Treponema pallidum antigen, and inserted it into Escherichia coli to express it, and obtained syphilis recombinant antigens TPN17 and TPN47;

[0037] 2) Spotting on nitrocellulose membrane

[0038] Coat the anti-human IgM specific fragment μ chain monoclonal antibody on the nitrocellulose membrane IgM detection line, coat goat anti-syphilis antigen TPN17 and TPN47 IgG antibody on the control line, and dry; the anti-human IgM specific fragment The concentration of the μ chain monoclonal antibody is 1-4 mg / mL, and the IgG antibody of goat anti-syphilis antigens TPN17 and TPN47 is mixed by anti-TPN17-IgG antibody and anti-TPN47-IgG antibody in a volume ratio of 1:1, and the final conce...

Embodiment 1

[0063] Coat the anti-human IgM specific fragment μ chain monoclonal antibody on the nitrocellulose membrane (NC membrane) IgM detection line, and coat the goat anti-syphilis antigen (TPN17 and TPN47) antibody on the control line, dry at room temperature, and seal at room temperature Save for later. Among them, the concentration of the anti-human IgM specific fragment μ chain monoclonal antibody is 1 mg / mL, and the goat anti-syphilis antigen (TPN17 and TPN47) IgG antibody is mixed by anti-TPN17-IgG antibody and anti-TPN47-IgG antibody in a volume ratio of 1:1 , the final concentration is 1mg / mL; the sample volume of the two is 1μL / cm.

[0064]The purified gold-labeled recombinant syphilis antigens TPN17 and TPN47 were mixed at a volume ratio of 1:1, evenly spread on glass fiber paper, dried at 37°C, prepared into a gold colloid pad, and sealed for later use. The solid-phase fiber membrane combined with colloidal gold, glass fiber, absorbent paper, etc. are combined in a certai...

Embodiment 2

[0067] Similar to Example 1, the difference is that the gold colloid pad is only composed of TPN17 and does not contain TPN47. Result judgment is identical with embodiment 1.

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Abstract

The invention provides a reagent strip for testing the syphilis specific IgM antibodies through gold immunochromatographic assay and a preparation method thereof and relates to a reagent for testing the syphilis specific IgM antibodies. The reagent strip is provided with a carrier plate, a sample-adding pad, a colloidal gold pad, a nitrocellulose membrane, a syphilis specific IgM antibody testing line, a control line and an absorption pad. The method comprises the following steps: preparing the recombinant syphilis antigens TPN17 and TPN47; applying samples of the nitrocellulose membrane; preparing the colloidal gold; labeling the colloidal gold and TPN17 and TPN47; and preparing the strip for testing through immunochromatographic assay. The invention has the following advantages: the specimen quantity needed for testing is small, special instruments are not needed, the results can be directly identified by naked eyes, testing is simple, convenient and rapid, the specificity is strong, the sensitivity is high, testing is accurate and reliable, the cost is low and the invention is widely applied.

Description

technical field [0001] The invention relates to a syphilis-specific IgM antibody detection reagent, in particular to a syphilis-specific IgM antibody rapid detection reagent strip and a preparation method thereof by using a colloidal gold immunochromatography technique (immunochromatography). Background technique [0002] Syphilis (Syphilis) is a sexually transmitted disease caused by Treponema pallidum (TP), and its pathogen is Treponema pallidum, which belongs to the family Treponemaceae. Treponema pallidum is mainly transmitted through sexual contact, blood transfusion, wound or placenta. Treponema pallidum enters the blood from the lymph nodes near the infected area, spreads throughout the body, and affects almost all tissues and organs of the body. The World Health Organization (WHO) once predicted optimistically: "Due to the availability of highly sensitive detection methods and efficient treatment options, syphilis is a sexually transmitted disease that can be succes...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/532G01N33/558G01N33/571
Inventor 林丽蓉张忠英杨天赐张长弓
Owner ZHONGSHAN HOSPITAL XIAMEN UNIV
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