Mycoplasma pneumoniae IgM antibody colloid gold method detecting kit and preparation method thereof

A technology of mycoplasma pneumoniae and colloidal gold, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low sensitivity, poor specificity of mycoplasma pneumoniae, and limited clinical application value, achieving high sensitivity, obvious effect, and easy operation Effect

Inactive Publication Date: 2012-01-04
北京中检安泰诊断科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cold aggregation test, indirect hemagglutination inhibition test, complement fixation test and culture of Mycoplasma pneumoniae have limited clinical application value due to problems such as poor specificity and low sensitivity.

Method used

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  • Mycoplasma pneumoniae IgM antibody colloid gold method detecting kit and preparation method thereof
  • Mycoplasma pneumoniae IgM antibody colloid gold method detecting kit and preparation method thereof
  • Mycoplasma pneumoniae IgM antibody colloid gold method detecting kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] 1. Source of raw materials

[0059] The source and dosage of each raw material and diluent prepared by the Mycoplasma pneumoniae IgM antibody detection kit (colloidal gold method) are shown in Table 10 below:

[0060]

[0061] 2. Raw material requirements:

[0062] 2.1 Recombinant MP antigen

[0063] (1) Appearance: colorless transparent liquid;

[0064] (2) Concentration and purity requirements: the concentration is greater than 2mg / ml, determined by SDS-PAGE, and there is only one band under the condition of 10μl sample loading;

[0065] 2.2 Goat anti-mouse IgG antibody

[0066] (1) Appearance: colorless transparent liquid;

[0067] (2) Concentration requirements: concentration greater than 4mg / ml;

[0068] 2.3 Mouse anti-human IgM monoclonal antibody

[0069] (1) Appearance: colorless transparent liquid;

[0070] (2) Concentration and purity requirements: the concentration is greater than 2 mg / ml, detected by SDS-PAGE protein electrophoresis, and the he...

Embodiment 2

[0151] Kit preparation one

[0152] (1) Preparation of reaction membrane: Dilute the recombinant MP antigen with phosphate buffer solution to a coating concentration of 1.0mg / ml, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0mg / ml, and use a film dispenser to apply The two coating solutions were drawn onto the nitrocellulose membrane respectively, and the coated nitrocellulose membrane was dried and stored;

[0153] (2) Preparation of mouse anti-human IgM monoclonal antibody gold conjugate pad: Colloidal gold-labeled mouse anti-human IgM monoclonal antibody solution was prepared by coupling mouse anti-human IgM monoclonal antibody with a labeling concentration of 10 μg / ml to colloidal gold. Centrifuge at 12000r / min for 40min, discard the supernatant, add the colloidal gold conjugate diluent to 1 / 2 of the original volume, then soak the gold standard pad with 1.5ml / strip of the mixed solution to prepare MP-IgM gold conjugate pad , put the MP-IgM go...

Embodiment 3

[0157] Kit Preparation II

[0158] (1) Preparation of reaction membrane: Dilute the recombinant MP antigen with phosphate buffer solution to a coating concentration of 1.0mg / ml, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0mg / ml, and use a film dispenser to apply The two coating solutions were drawn onto the nitrocellulose membrane respectively, and the coated nitrocellulose membrane was dried and stored;

[0159] (2) Preparation of mouse anti-human IgM monoclonal antibody gold conjugate pad: Colloidal gold-labeled mouse anti-human IgM monoclonal antibody solution was prepared by coupling mouse anti-human IgM monoclonal antibody with a labeling concentration of 18 μg / ml to colloidal gold. Centrifuge at 12000r / min for 40min, discard the supernatant, add the colloidal gold conjugate diluent to 1 / 2 of the original volume, then soak the gold standard pad with 1.5ml / strip of the mixed solution to prepare MP-IgM gold conjugate pad , put the MP-IgM gol...

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Abstract

The invention relates to a mycoplasma pneumoniae IgM antibody colloid gold method detecting kit which comprises a nitrocellulose membrane detecting line coated recombinant antigen MP-Ag, a quality control line coated goat anti-mouse IgG antibody and a gold labeled pad coated mouse anti-human IgM monoclonal antibody which is marked by colloidal gold, wherein the recombinant mycoplasma pneumoniae antigen is colorless transparent liquid, has the concentration of larger than 2mg/ml and is detected by SDS-PAG; the goat anti-mouse IgG antibody is colorless transparent liquid and has the concentration of larger than 4mg/ml; the mouse anti-human IgM monoclonal antibody is colorless transparent liquid, has the concentration of larger than 2mg/ml and is detected by SDS-PAGE; and the sample loading amount uses two strips under the condition of 10 microliters. The mycoplasma pneumoniae IgM antibody colloid gold rapid detecting test paper tape uses multiepitope recombinant antigen as a raw materials, has the characteristics of simpler operation, low cost, high specificity and high sensitivity, can carry out single detection, is easy for popularization and has obvious detecting and control effect on the mycoplasma pneumoniae IgM antibody.

Description

technical field [0001] The invention relates to an antibody detection kit and a preparation method thereof, in particular to a mycoplasma pneumoniae IgM antibody colloidal gold method detection kit and a preparation method thereof. Background technique [0002] Mycoplasma pneumoniae (MP) is a common pathogenic microorganism that causes primary atypical pneumonia and other respiratory infectious diseases. In addition to the respiratory tract, MP can still cause serious complications in other systems. [0003] Mycoplasma pneumoniae antibody detection methods include: cold aggregation test, indirect hemagglutination inhibition test, complement fixation test, culture of Mycoplasma pneumoniae, polymerase chain reaction (PCR). The diagnosis of mycoplasma pneumonia includes pathogen culture, a variety of specific serological examinations, and in recent years, gene probes, DNA, and PCR methods. However, cold aggregation test, indirect hemagglutination inhibition test, complement f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531G01N33/532
Inventor 李习荣
Owner 北京中检安泰诊断科技有限公司
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