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

A technology of immunochromatography detection and antibody detection, which is applied in the direction of measuring devices, analysis materials, instruments, etc.

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

Examples

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preparation example Construction

[0033] The preparation method of the syphilis-specific IgG antibody colloidal gold immunochromatographic detection reagent strip includes the following steps:

[0034] 1) Preparation of syphilis recombinant antigens TPN17 and TPN47

[0035] Using gene cloning technology, PCR amplifies the DNA encoding the Treponema pallidum antigen, and inserts it into Escherichia coli for expression to obtain the syphilis recombinant antigens TPN17 and TPN47.

[0036] 2) Spotting of nitrocellulose membrane

[0037] Coat the anti-human IgG specific fragment γ chain monoclonal antibody on the nitrocellulose membrane IgG detection line, and coat the goat anti-syphilis antigen TPN17 and TPN47 IgG antibodies at the control line (C), and dry, the anti-human IgG The concentration of the specific fragment γ chain monoclonal antibody is 1~4mg / mL, the goat anti-syphilis antigen TPN17 and TPN47 IgG antibody is mixed by the volume ratio of anti-TPN17-IgG antibody and anti-TPN47-IgG antibody at a volume ratio of ...

Embodiment 1

[0064] Coat the anti-human IgG specific fragment γ chain monoclonal antibody on the syphilis-specific IgG antibody detection line, and coat the goat anti-syphilis antigen TPN17 and TPN47 antibodies on the control line, dry at room temperature, and store in a sealed room for later use. Among them, the concentration of anti-human IgG specific fragment γ chain monoclonal antibody is 1 mg / mL, and goat anti-syphilis antigen (TPN17 and TPN47) IgG antibodies are mixed by anti-TPN17-IgG antibody and anti-TPN47-IgG antibody in a volume ratio of 1:1 , Its final concentration is 1mg / mL; the spotting amount for both is 1μl / cm.

[0065] After the purified gold-labeled syphilis recombinant antigens TPN17 and TPN47 were mixed at a volume ratio of 1:1, they were evenly coated on glass fiber paper, dried at 37°C, and prepared into a gold conjugate pad, which was sealed for later use. Combine the solid-phased fiber membrane with colloidal gold and combine glass fiber, absorbent paper, etc., in a c...

Embodiment 2

[0068] Similar to Example 1, the difference is that the colloidal gold pad only consists of TPN17 and does not contain TPN47. The result was judged to be the same as in Example 1.

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Abstract

The invention provides a reagent strip for testing the syphilis specific IgG antibodies in the specimens of whole blood, serum, blood plasma, cerebrospinal fluid and the like through gold immunochromatographic assay and a preparation method thereof and relates to a reagent for testing the syphilis specific IgG antibodies. The reagent strip is provided with a carrier plate, a sample-adding pad, a colloidal gold pad, a nitrocellulose membrane, a syphilis specific IgG 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 relatively high, testing is accurate and reliable, the cost is relatively low and the invention is widely applied.

Description

Technical field [0001] The invention relates to a syphilis-specific IgG antibody detection reagent, in particular to a syphilis-specific IgG antibody rapid detection reagent strip using colloidal gold immunochromatography (immunochromatography) and a preparation method thereof. Background technique [0002] Syphilis is a sexually transmitted disease caused by Treponema pallidum (TP), and its pathogen is Treponema pallidum, belonging to the family Treponema. Treponema pallidum is mainly transmitted through sexual contact, blood transfusion, wounds or placenta. Treponema pallidum enters the blood from the lymph nodes near the infected area and spreads throughout the body, causing almost all tissues and organs of the body to be affected. The clinical manifestations are systemic and can be divided into different clinical stages, including the first, second, third and incubation periods. The World Health Organization (WHO) has optimistically predicted: "Because of high-sensitivity te...

Claims

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Application Information

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