Chlamydia pneumoniae IgM (immunoglobulin M) colloidal golden method kit and preparation method thereof

A technology of chlamydia pneumoniae and detection kit, which is applied in the directions of measuring devices, instruments, scientific instruments, etc., to achieve the effects of easy popularization, simple operation and obvious effect

Inactive Publication Date: 2012-10-24
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

Detection by linking polymerase chain reaction (LCR) can further improve the sensitivity and detection rate, but it has not been used clinically

Method used

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  • Chlamydia pneumoniae IgM (immunoglobulin M) colloidal golden method kit and preparation method thereof
  • Chlamydia pneumoniae IgM (immunoglobulin M) colloidal golden method kit and preparation method thereof
  • Chlamydia pneumoniae IgM (immunoglobulin M) colloidal golden method kit and preparation method thereof

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Experimental program
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Embodiment 1

[0114] 1. Source of raw materials

[0115] See Table 17 below for the sources and amounts of raw materials and diluents prepared by the Chlamydia pneumoniae IgM antibody colloidal gold method detection kit:

[0116] Table 17

[0117]

[0118] 2. Raw material requirements:

[0119] 2.1 Recombinant Cpn antigen

[0120] (1) Appearance: colorless transparent liquid;

[0121] (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;

[0122] 2.2 Goat anti-mouse IgG antibody

[0123] (1) Appearance: colorless transparent liquid;

[0124] (2) Concentration requirements: concentration greater than 4mg / ml;

[0125] 2.3 Mouse anti-human IgM monoclonal antibody

[0126] (1) Appearance: colorless transparent liquid;

[0127] (2) Concentration and purity requirements: the concentration is greater than 2 mg / ml, detected by SDS-PAGE protein electrophoresis,...

Embodiment 2

[0179] Kit preparation one

[0180] (1) Preparation of the reaction membrane: Dilute the recombinant Cpn-Ag with phosphate buffer solution to a coating concentration of 1.0 mg / ml, dilute the goat anti-mouse IgG antibody to a concentration of 2.0 mg / ml, and dilute the two coating solutions Draw on the nitrocellulose membrane respectively, and store the coated nitrocellulose membrane after drying;

[0181] (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 5 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 label pad with the mixed solution to prepare a Cpn-IgM gold conjugate pad, and mix the Cpn-IgM Store gold conjugate pads dry;

[0182] (3) Cutting a...

Embodiment 3

[0184] Kit Preparation II

[0185] (1) Preparation of the reaction membrane: Dilute the recombinant Cpn-Ag with phosphate buffer solution to a coating concentration of 1.0 mg / ml, dilute the goat anti-mouse IgG antibody to a concentration of 2.0 mg / ml, and dilute the two coating solutions Draw on the nitrocellulose membrane respectively, and store the coated nitrocellulose membrane after drying;

[0186] (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 30 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 label pad with the mixed solution to prepare a Cpn-IgM gold conjugate pad, and mix the Cpn-IgM Store gold conjugate pads dry;

[0187] (3) Cutting a...

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Abstract

The invention discloses a chlamydia pneumoniae IgM (immunoglobulin M) colloidal golden method kit which comprises recombinant chlamydia pneumoniae antigens enveloped on a nitrocellulose membrane detection line, goat anti-rat IgG (immunoglobulin G) antibodies enveloped on a quality control line and rat anti-human IgM monoclonal antibodies which have colloidal gold labels and are enveloped on a gold label pad, the concentration of the chlamydia pneumoniae antigens ranges from 1mg / ml to 2mg / ml and is measured by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis), the concentration of the goat anti-rat IgG (immunoglobulin G) antibodies ranges from 1mg / ml to 3mg / ml, and the concentration of the rat anti-human IgM monoclonal antibodies ranges from 5g / mL to 30g / mL and is measured by the SDS-PAGE. The chlamydia pneumoniae IgM colloidal golden method kit has the advantages that the kid is speedy, simple, convenient and accurate and is high in sensitivity, and a judgment result can be read after integral operation time of dozens of minutes; and a colloidal gold quick detecting paper strip is made of the multi-epitope recombinant antigens and is simple and convenient to operate, low in cost, good in specificity and high in sensitivity, can be used for single-component detection and is popularized easily, and detection and control effects for chlamydia pneumoniae IgM are obvious.

Description

technical field [0001] The invention relates to an antibody detection kit and a preparation method thereof, in particular to a chlamydia pneumoniae IgM antibody colloidal gold method detection kit and a preparation method thereof. Background technique [0002] Chlamydia pneumoniae is an important pathogen of human respiratory diseases, which can cause acute and chronic respiratory diseases. Surveys show that 5-10% of community-acquired pneumonia, bronchitis and sinusitis are caused by Chlamydia pneumoniae. It often causes atypical pneumonia in adults and adolescents, and can also cause acute respiratory infections such as bronchitis, pharyngitis and tonsillitis. The clinical manifestations of Chlamydia pneumoniae respiratory infection are not typical, usually with sore throat and hoarseness onset, followed by coughing a few days to a week later, compared with other respiratory diseases, Chlamydia pneumoniae infection takes the longest time from onset to medical treatment, so...

Claims

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Application Information

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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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