Mycoplasma pneumoniae IgM antibody colloidal gold immunochromatography detection kit and preparation method thereof

An immunochromatographic detection and detection kit technology, applied in the field of immunochromatography, can solve the problems of large batch-to-batch variation, unstable gold-labeled substances, and low sensitivity.

Active Publication Date: 2021-10-22
SHANGHAI BIOGERM MEDICAL TECH CO LTD BEIJING BRANCH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the colloidal gold detection technology for Mycoplasma pneumoniae in the prior art still has the problems of large batch-to-batch variation, low sensitivity, and unstable gold standard substances.

Method used

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  • Mycoplasma pneumoniae IgM antibody colloidal gold immunochromatography detection kit and preparation method thereof
  • Mycoplasma pneumoniae IgM antibody colloidal gold immunochromatography detection kit and preparation method thereof
  • Mycoplasma pneumoniae IgM antibody colloidal gold immunochromatography detection kit and preparation method thereof

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preparation example Construction

[0050] The present invention also provides a preparation method of the above-mentioned detection kit, wherein the sample pad, binding pad, detection pad and water-absorbing pad are sequentially pasted on the bottom plate to obtain a detection test strip; and the dilution of the sample to be tested is obtained by configuring the amount according to the formula.

[0051] In order to reduce the batch-to-batch difference of test strips and improve the specificity of the kit, the inventors optimized the preparation process of the binding pad. On the one hand, the preparation process of colloidal gold adopts the trisodium citrate reduction method, and the preparation process adopts spherical Bottle, cantilever magnetic stirring paddle and digital display jacket magnetic stirrer keep the temperature of the colloidal gold solution under control, and the size of the fired gold particles is consistent; on the other hand, optimize the colloidal gold-labeled mouse anti-human IgM monoclonal ...

Embodiment 1

[0053] Example 1 Mycoplasma pneumoniae IgM antibody colloidal gold immunochromatography detection kit

[0054] 1. Buffer system configuration:

[0055] Colloidal gold-labeled mouse anti-human IgM monoclonal antibody dilution: 50mM Tris-HCl, 3(w / v)% sucrose, 3(w / v)% trehalose, 1(w / v)%BSA, 0.2( w / v) % PVP40, 1 (w / v) % casein and 0.7 (w / v) % Tween-20.

[0056] Dilution of Mycoplasma pneumoniae antigen: 10 mM PBS, 2 (w / v) % trehalose, 0.5 mM disodium EDTA, 0.1 (w / v) % Triton X-100 and 0.1 (w / v) % BSA.

[0057] Diluent of goat anti-mouse IgG polyclonal antibody: 10 mM PBS, 3 (w / v)% trehalose and 0.5 mM disodium EDTA.

[0058] The treatment solution of the sample pad: 50mM Tris-HCl, 1(w / v)% BSA, 0.5(w / v)% casein, 100μg / mL mouse anti-human RBC antibody, 50mg / mL lectin and 0.1(w / v) v) % Tween-20.

[0059] Sample diluent to be tested: 20 mM Tris-HCl, 0.1 (w / v)% S9, 0.3 (w / v)% S17 and 50 mM NaCl.

[0060] 2. Firing of colloidal gold solution around 30nm:

[0061] (1) Add 1000ml of...

Embodiment 2

[0083] Example 2 Colloidal gold-labeled mouse anti-human IgM monoclonal antibody mouse anti-human IgM monoclonal antibody dosage optimization

[0084] Take 8 small glass tubes, add 1mL colloidal gold to each, and then add 0.1mol / L K 2 CO 3 7μl, after mixing evenly, add 1mg / mL anti-human IgM monoclonal antibody 1, 2, 3, 4, 5, 8, 10, 12μl respectively, mix evenly, after standing for 5min, add 200μl of 10% NaCl, mix evenly, let stand Observe the color change after 5 minutes. The results are shown in Table 1.

[0085] Table 1

[0086]

[0087] Table 2 is the color change of each tube when measuring the amount of mouse anti-human IgM. The color of the first No. 5 samples turns blue or purple after adding NaCl, which shows that the colloidal gold is agglomerated, and the colloidal gold at this time is unstable. Colloidal gold after sample No. 6 is stable. In general, the amount of protein should be in excess, so sample No. 7 is selected here, that is, 1 mL of colloidal gold...

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Abstract

The invention relates to the field of immunochromatography, and particularly provides a mycoplasma pneumoniae IgM antibody colloidal gold immunochromatography detection kit and a preparation method thereof. The mycoplasma pneumoniae IgM antibody colloidal gold immunochromatography detection kit comprises a detection test strip and a to-be-detected sample diluent, the detection test strip comprises a bottom plate and a sample pad, a combination pad, a detection pad and a water absorption pad which are sequentially arranged on the bottom plate. The kit detects a to-be-detected sample according to the immunochromatography principle, is low in cost and short in time consumption, the to-be-detected sample is diluted by the diluent containing 10-100 mM of Tris-HCl, 0.1-0.5 (w/v)% of S9, 0.1-0.5 (w/v)% of S17 and 10-100 mM of NaCl, the detection sensitivity and specificity can be remarkably improved, and the detection accuracy of mycoplasma pneumoniae is improved.

Description

technical field [0001] The invention relates to the field of immunochromatography, in particular to a colloidal gold immunochromatography detection kit for mycoplasma pneumoniae IgM antibody and a preparation method thereof. Background technique [0002] Mycoplasma pneumoniae (MP) is a prokaryotic microorganism without a cell wall, belonging to the mollusc class, and is the main pathogen that causes community-acquired pneumonia. [0003] At present, the methods for diagnosis of Mycoplasma pneumoniae include: isolation and culture, colloidal gold method, passive agglutination method, enzyme-linked immunosorbent assay, complement fixation test, fluorescent quantitative PCR and so on. Among them, the culture period of isolated and cultured pathogens is longer; passive agglutination and complement fixation tests cannot distinguish between IgM and IgG; although enzyme-linked immunoassay and fluorescent quantitative PCR have the advantages of high sensitivity and specificity, they...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/569G01N33/558G01N33/543G01N33/52
CPCG01N33/587G01N33/56933G01N33/558G01N33/54346G01N33/52G01N2469/20G01N2333/30
Inventor 梁权赵百慧刘尚奇魏鹏海储寒君张淑芳
Owner SHANGHAI BIOGERM MEDICAL TECH CO LTD BEIJING BRANCH
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