Reagent strip for joint detection of syphilis specific IgM antibody and specific total antibody and preparation method thereof

A combined detection and antibody detection technology, applied in measurement devices, instruments, scientific instruments, etc., to achieve the effects of high sensitivity, wide application and strong specificity

Active Publication Date: 2010-09-08
ZHONGSHAN HOSPITAL XIAMEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although a negative TP-IgM does not completely rule out infectivi

Method used

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  • Reagent strip for joint detection of syphilis specific IgM antibody and specific total antibody and preparation method thereof
  • Reagent strip for joint detection of syphilis specific IgM antibody and specific total antibody and preparation method thereof
  • Reagent strip for joint detection of syphilis specific IgM antibody and specific total antibody and preparation method thereof

Examples

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preparation example Construction

[0041] The preparation method of the joint detection reagent strip of the syphilis-specific IgM antibody and the specific total antibody comprises the following steps:

[0042] 1) Preparation of recombinant syphilis antigens TPN17 and TPN47

[0043] Using gene cloning technology, PCR amplified the DNA encoding Treponema pallidum antigen, and inserted it into Escherichia coli to express it, and obtained syphilis recombinant antigens TPN17 and TPN47.

[0044] 2) Spotting on nitrocellulose membrane

[0045] Anti-human IgM-specific fragment μ chain monoclonal antibody is coated on the syphilis-specific IgM antibody detection line, and anti-human Ig monoclonal antibody or syphilis-specific antigen TPN17 and / or syphilis-specific antibody is coated on the specific total antibody detection line Sexual antigen TPN47, dry, the concentration of the anti-human Ig monoclonal antibody or syphilis-specific antigen TPN17 and / or syphilis-specific antigen TPN47 can be 1 ~ 4mg / mL, anti-human Ig...

Embodiment 1

[0071] The first reagent strip A is coated with anti-human IgM specific fragment μ chain monoclonal antibody on the nitrocellulose membrane (NC membrane) IgM detection line, and the specific total antibody detection line of the second reagent strip B is coated with anti-human Ig Monoclonal antibody, coated with goat anti-syphilis antigen TPN17 and TPN47 antibodies at the control line C, dried at room temperature, sealed and stored at room temperature for future use. Among them, the concentration of anti-human IgM specific fragment μ chain monoclonal antibody and anti-human Ig monoclonal antibody is 1 mg / mL, goat anti-syphilis antigen (TPN17 and TPN47) IgG antibody is composed of anti-TPN17-IgG antibody and anti-TPN47-IgG antibody Mix according to the volume ratio of 1:1, the final concentration is 1mg / mL; the sample volume of the three is 1μL / cm.

[0072] The purified gold-labeled recombinant syphilis antigens TPN17 and TPN47 were mixed at a volume ratio of 1:1, evenly spread ...

Embodiment 2

[0075] Similar to Example 1, the difference is that the gold colloid pad and the syphilis-specific total antibody detection line consist only of TPN17 and do not contain TPN47. Result judgment is identical with embodiment 1.

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Abstract

The invention provides a reagent strip for joint detection of a syphilis specific IgM antibody and a specific total antibody and a preparation method thereof, relating to a reagent for joint detection of a syphilis specific IgM antibody and a specific total antibody. The reagent strip is provided with two reagent strips, wherein the two reagent strips are both provided with vector plates, sample adding pads, colloidal gold pads, nitrocellulose membranes, control lines and absorbent pads and respectively provided with a syphilis specific IgM antibody detection line and a specific total antibody detection line. The preparation method comprises the following steps: preparing recombinant syphilis antigens TPN17 and TPN47, applying samples of nitrocellulose membranes, preparing colloidal gold,labeling TPN17 and TPN47 with colloidal gold and preparing immunochromatography detection strips. The reagent strip can be used for detecting the syphilis specific IgM antibody and the specific totalantibody in the specimens of whole blood, blood serum, blood plasma, cerebrospinal fluid and the like. During detection, the specimen amount is minimal, special instruments are not needed and the results are directly interpreted by the naked eyes. The reagent strip is simple, convenient, fast, accurate and reliable and has strong specificity, high sensitivity, low cost and wide application range.

Description

technical field [0001] The invention relates to a combined detection reagent for syphilis-specific IgM antibodies and specific total antibodies, in particular to a reagent strip for the combined detection of syphilis-specific IgM antibodies and specific total antibodies using colloidal gold immunochromatography (immunochromatography) and its preparation method. 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...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/571G01N33/543
Inventor 张忠英林丽蓉杨天赐张长弓
Owner ZHONGSHAN HOSPITAL XIAMEN UNIV
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