Three-in-one detection kit for cardiac troponin I, creatine kinase isoenzyme and myoglobin and preparation method of three-in-one detection kit

A cardiac troponin and detection kit technology, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of strong interference and high intensity, and achieve the effects of high sensitivity, fast response and strong specificity

Inactive Publication Date: 2018-02-06
WEIHAI NEOPROBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the usual fluorescence measurement, because the test sample contains a variety of fluorescent components, the background fluorescence (scattered light caused by colloidal particles and solvent molecules in the sample and non-specific fluorescence emitted by proteins and other compounds in the serum) has a large intensity and interference. Strong, become the bottleneck of large-scale promotion of fluorescence analysis

Method used

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  • Three-in-one detection kit for cardiac troponin I, creatine kinase isoenzyme and myoglobin and preparation method of three-in-one detection kit
  • Three-in-one detection kit for cardiac troponin I, creatine kinase isoenzyme and myoglobin and preparation method of three-in-one detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The various components of the test paper card in the cardiac troponin I, creatine kinase isoenzyme and myoglobin three-in-one detection kit can be prepared by the following measures:

[0037] 1. Preparation of sample pad 2:

[0038] Soak the glass fiber membrane in the treatment solution containing 2.0% Triton X-100, 2% BSA, 0.1M Tris buffer, pH7.5, soak at 4°C for 4 hours, then place it in an oven and dry it at 37°C 2 hours. 2. Preparation of binding pad 3 for absorbing fluorescent microsphere-labeled antibody:

[0039] Soak the glass fiber membrane in 200mM Tris-HCL treatment solution (containing 1.5% Triton X-100, 1.5% BSA, pH7.5), soak at 4°C for 4 hours, then take it out of a 37°C oven and dry it for 4 hours, and set it aside. Put the glass fiber membrane on the Bio-DotXYZ3050 three-dimensional spraying platform, and use the Bio-Jet Quanti300 non-contact micro-quantitative nozzle to combine rare earth fluorescent microspheres labeled cardiac troponin I, creatine ...

Embodiment 2

[0048] Embodiment 2: accuracy test

[0049] Select the above test paper card and fluorescence immunochromatography analyzer (model: NEO-007),

[0050] Setting of the parameters of the fluorescence immunoassay analyzer: After setting the process parameters of the test paper card on the fluorescence immunoassay analyzer, take the above-mentioned assembled test paper card, and use 0.2, 0.5, 1, 2, 5, 20 ng / ml of myocardial muscle Calpain I, creatine kinase isozyme and myoglobin three-in-one calibrator are measured with a test paper card to obtain the fluorescence intensity value of each calibrator, and the results are input into the parameters of the analyzer to complete the parameters of the analyzer. settings.

[0051] Main testing materials: clinical samples obtained from relevant hospitals, a total of 200 Roche electrochemiluminescence immunoassay value samples, including 100 serum samples, 100 whole blood samples, cardiac troponin I, creatine kinase isoenzyme and myoglobin ...

Embodiment 3

[0060] Embodiment 3: precision test

[0061] Using the test paper card and measuring system of Example 2, the test paper card and the fluorescent immunochromatographic analyzer of the present invention were tested for precision.

[0062] Main testing materials: clinical samples were obtained from relevant hospitals, a total of 2 serum samples with chemiluminescence immunoassay value, among which the clinical measurement value of the low value fixed value sample was 0.24ng / ml, and the clinical measurement value of the high value fixed value sample was 1.78ng / ml .

[0063] Preparation:

[0064] Using the test paper card and measuring system of Example 2, each of the 2 fixed-value samples was repeatedly measured 20 times.

[0065] Analysis of test results:

[0066] After the clinical sample testing reagents are prepared, the clinical samples are tested according to the preparation method, and the test results are analyzed.

[0067] test results:

[0068] As shown in Table 1, t...

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Abstract

The invention relates to the technical field of fluorescence immunochromatography in medical immunology and in particular relates to a three-in-one detection kit for cardiac troponin I, creatine kinase isoenzyme and myoglobin and a preparation method of the three-in-one detection kit. The three-in-one detection kit comprises a test paper card, and is characterized in that a PVC (Polyvinyl Chloride) panel, a sample pad, a conjugate pad, a nitrocellulose membrane and an absorbent pad are sequentially arranged on the test paper card from bottom to top, wherein a cardiac troponin I, creatine kinase isoenzyme and myoglobin three-in-one monoclonal antibody labeled with rare-earth fluorescence microspheres is adsorbed onto the conjugate pad; the diameter of the rare-earth fluorescence microspheres is 60-120nm; the rare-earth fluorescence microspheres are doped with rare-earth lanthanide elements, are stable in a ground state and can emit fluorescence in a wavelength range of 540-600nm under the action of an excitation light source of 340-380nm; and the monoclonal antibody is a purified and mixed monoclonal antibody and is derived from a monoclonal antibody cell strain targeted at 2-6 different cardiac troponin I, creatine kinase isoenzyme and myoglobin three-in-one antigen epitopes. The three-in-one detection kit disclosed by the invention has the advantages of being simple and convenient in operation, rapid in response, high in sensitivity, high in specificity and the like.

Description

Technical field: [0001] The invention relates to the technical field of fluorescent immunochromatography in medical immunology, in particular to a cardiac troponin I, creatine kinase isoenzyme and myoglobin three-in-one quantitative analysis of cardiac troponin I A three-in-one detection kit for calpain I, creatine kinase isozyme and myoglobin and a preparation method thereof. Background technique: [0002] cTnI is a myocardial structural protein with a high degree of tissue specificity. It is a very sensitive and specific marker of acute myocardial infarction. cTnI is abundant in the myocardium, and it is released into the blood 4 to 6 hours after myocardial injury, reaching the diagnostic decision value. The peak appears at 14 hours to 36 hours and lasts for about 3 days to 7 days. The window period is longer, up to about 10 days, which is conducive to the diagnosis of delayed AMI and unstable angina. CK-MB is an isoenzyme of creatine kinase (CK), and CK is the most wide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/573G01N33/533G01N33/558G01N33/577
CPCG01N33/533G01N33/558G01N33/573G01N33/577G01N33/6893G01N2800/324G01N2333/4712
Inventor 王鹏浩宋璐琳张培镇于鸿翔孙宁宁
Owner WEIHAI NEOPROBIO
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