Immunofluorescence dipstick component for quickly and quantitatively detecting protein of plurality of types and detection card component prepared from same and preparation method thereof

A quantitative detection and immunofluorescence technology, applied in the field of medical testing, can solve problems such as the inability to meet the accurate and quantitative requirements of clinical diagnosis, the inability to monitor the course of AMI, and the expensive detection equipment, etc. low cost effect

Active Publication Date: 2012-09-12
GUANGZHOU HONGQI OPTICAL INSTR TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

3) It lasts for a long time in the blood. If it appears transiently or exists in the blood for a short time, it is difficult to seize the right time to detect a positive result, which may easily lead to missed diagnosis
[0008] At present, the detection of myoglobin, CK-MB and cTnI mostly adopts chemiluminescence method, which is labeled with chemiluminescent substances as detection antibodies, which greatly improves the accuracy and sensitivity of the determination, but requires expensive detection equipment, whi...

Method used

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  • Immunofluorescence dipstick component for quickly and quantitatively detecting protein of plurality of types and detection card component prepared from same and preparation method thereof
  • Immunofluorescence dipstick component for quickly and quantitatively detecting protein of plurality of types and detection card component prepared from same and preparation method thereof
  • Immunofluorescence dipstick component for quickly and quantitatively detecting protein of plurality of types and detection card component prepared from same and preparation method thereof

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Effect test

Embodiment 1

[0059] refer to figure 1 As shown, an immunofluorescence test strip assembly for rapid quantitative detection of various proteins of the present invention is composed of a substrate 1 and a water-absorbing pad 2 sequentially bonded on the substrate 1, a coated analysis membrane 3, and a sample pad 6 The test strip and the independently packaged fluorescein-labeled specific antibody are combined; there are three detection lines 4 and one quality control line 5 on the coating analysis membrane 3, and the three detection lines 4 on the coating analysis membrane respectively coated with an anti-myoglobin monoclonal antibody line, an anti-creatine kinase isoenzyme monoclonal antibody line and a troponin I monoclonal antibody line, the quality control line 5 is coated with rabbit IgG antibody; The independently packaged fluorescein-labeled specific antibodies are separately packaged anti-myoglobin monoclonal antibody, anti-creatine kinase isoenzyme monoclonal antibody, anti-troponin...

Embodiment 2

[0093] The preparation method of the present embodiment is basically the same as that of the first embodiment, the difference is that:

[0094] In step 2, use 50mM pH7.6 phosphate buffer, containing methanol 0.8%, sucrose 1%, bovine serum albumin 0.6% anti-myoglobin monoclonal antibody, anti-creatine kinase isoenzyme monoclonal antibody, anti-myoglobin The concentration of the calpain I monoclonal antibody is 1 mg / m; the preparation method of the quality control line coating buffer is: dilute with polybutene buffer solution of 50mM pH7.6, rabbit containing 0.7% methanol and 0.5% bovine serum albumin The concentration of IgG antibody is up to 0.5mg / ml; the preparation of the coating analysis membrane: adjust the spraying machine, the volume of the membrane liquid is 20ul / 40cm, the machine scribes, the distance between the detection line and the quality control line is 5mm, the scribing is fine and uniform, and it is placed for 25 ℃-37℃ vacuum drying oven for 1.5 hours, bagged a...

Embodiment 3

[0096] The preparation method of the present embodiment is basically the same as that of the first embodiment, the difference is that:

[0097] In step 3, the anti-myoglobin monoclonal antibody, anti-creatine kinase isoenzyme monoclonal antibody, anti-troponin I monoclonal antibody and anti-rabbit IgG antibody were diluted to 1 mg / ml, each take 5ml of antibody solution, add 40mg of fluorescent material platinum porphyrin solution, stir well, incubate at room temperature for 1.5 hours, and mix every 15 minutes. Finally, use a G25 gel column with a specification model for separation and purification, collect the labeled fluorescein-labeled antibody, and use a gel containing 0.01%-0.5% polyethylene glycol, 1%-5% bovine serum albumin, 5%-20% Glycerin and 0.01%-0.05% surfactant are diluted with 0.01M phosphate buffer solution, sealed and packaged in plastic bottles, and stored at 4°C.

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Abstract

The invention discloses an immunofluorescence dipstick component for quickly and quantitatively detecting protein of a plurality of types and a detection card component prepared from the same and a preparation method thereof, wherein the protein of a plurality of types comprises muscle hemoglobin/creatine kinase isoenzyme/troponin I; the dipstick component comprises a dipstick consisting of a bottom liner, a water-absorbing pad, a coated analysis membrane and a sample pad and independently packaged fluorescein mark specific antibodies; three detection lines and a quality control line are arranged on the coated analysis membrane; the detection line on the coated analysis membrane is respectively coated with an anti-myoglobin monoclonal antibody line, an anti-creatine kinase isozyme monoclonal antibody line and a troponin I monoclonal antibody line; the quality control line is coated with a rabbit IgG antibody; the fluorescein mark specific antibodies comprise the anti-myoglobin monoclonal antibody line, the anti-creatine kinase isozyme monoclonal antibody line, an troponin I monoclonal antibody and an anti-rabbit IgG antibody; and the detection card component comprises the dipstick, a card box consisting of a cover plate and a back plate and independently packaged platinum porphyrin mark specific antibodies and can synchronously detect the muscle hemoglobin/creatine kinase isoenzyme/troponin I, is simple to operate, is quick and sensitive, and has good specificity.

Description

technical field [0001] The invention belongs to the field of medical testing, and in particular relates to an immunofluorescence test strip component for rapid quantitative and combined determination of myoglobin / creatine kinase isoenzyme / troponin I and a preparation method thereof. Background technique [0002] Coronary heart disease has become a major disease affecting the health of the population. Acute myocardial infarction (AMI) is the main cause of death in coronary heart disease patients. How to timely discover patients with acute myocardial infarction and corresponding treatment is the key to saving the lives of patients with myocardial infarction. Traditional diagnosis is mainly based on typical clinical manifestations, ECG changes and laboratory enzyme examination. However, a considerable number of patients with myocardial infarction have no obvious clinical manifestations, and there is no obvious change in the early ECG. In this case, the application of specifi...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 罗琪谢爱武梁万兴李必松
Owner GUANGZHOU HONGQI OPTICAL INSTR TECH
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