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nt‑probnp detection kit and detection method

A detection kit, the technology of nt-probNP, which is applied in the field of NT-proBNP detection kits, can solve the problems of radioactive contamination radiation, long operation time, poor quantitative accuracy, etc., and achieves clear clinical guiding significance, strong specificity and response. fast effect

Active Publication Date: 2017-05-17
WEIHAI NEOPROBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the ELISA method has poor quantitative accuracy, long operation time, and low degree of automation, and is mostly used for qualitative detection; the radioimmunoassay method can reach a sensitivity of 4pg / ml, and is simple to operate and accurate in measurement. Radiation risk; the electrochemiluminescence method has strong specificity, high sensitivity and high accuracy, but requires expensive instruments and equipment and experienced operators, and is generally used in specific medical institutions; although the colloidal gold immunochromatography method has sample The advantages of less dosage, simple and fast, and low cost, but the sensitivity is low, generally only qualitative, not quantitative, especially the shortcoming of poor repeatability limits its clinical application, especially not suitable for accurate quantification Quantitative detection of body fluid marker proteins to help diagnose diseases

Method used

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  • nt‑probnp detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Each component of the test paper card in the NT-proBNP detection kit can be prepared by the following measures:

[0036] 1. Preparation of sample pad 2:

[0037] Soak the glass fiber membrane in the treatment solution containing 2.0% TritonX-100, 2% BSA, 0.1MTris buffer, pH7.5, soak at 4°C for 4 hours, then place it in an oven, and dry it at 37°C for 2 hours .

[0038] 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 the oven at 37°C and dry it for 4 hours, and set it aside. Put the glass fiber membrane on the Bio-DotXYZ3050 three-dimensional spraying platform, use the Bio-Jet Quanti300 non-contact micro-quantitative nozzle to spray the NT-proBNP monoclonal antibody labeled with rare earth fluorescent microspheres on the glass fiber membrane, and dry it at 37°C f...

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 immunoassayer, take the above-mentioned assembled test paper card and use 0.2, 0.5, 1, 2, 5, 20ng / ml The proBNP calibrator is measured with a test paper card to obtain the fluorescence intensity value of each calibrator, and the result is input into the parameters of the analyzer to complete the setting of the parameters of the analyzer.

[0051] Main testing materials: clinical samples were obtained from relevant hospitals, a total of 200 Roche electrochemiluminescence immunoassay value samples, including 100 serum samples, 100 whole blood samples, and the distribution range of NT-proBNP content was between 0-20ng / mL.

[0052] Detection method:

[0053] Step 1: Equilibrate the det...

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] Detection method:

[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 test reagents are prepared, the clinical samples are tested according to the test method, and the test results are analyzed.

[0067] test results:

[0068] as attached image 3 As ...

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Abstract

The invention relates to the technical field of fluorescence immunochromatography in medical immunology, specifically to an NT-proBNP detection kit and a detection method. The NT-proBNP detection kit is provided with a test cassette and is characterized in that the test cassette is successively provided with, from bottom to top, a PVC plate, a sample pad, a combination pad, a cellulose nitrate film and a water-absorbing pad, wherein an NT-proBNP monoclonal antibody labeled by a rare earth fluorescent microsphere is adsorbed on the combination pad, the rare earth fluorescent microsphere has a diameter of 60 to 120 nm, is doped by rare earth lanthanide, is stable in a ground state and emits fluorescent light with a wavelength in a range of 540 to 600 nm under the action of an excitation light source in a wavelength range of 340 to 380 nm, and the monoclonal antibody is a purified mixed monoclonal antibody and is originated from monoclonal antibody cell strains directed at 2 to 6 different NT-proBNP epitopes. The NT-proBNP detection kit has the advantages of simple operation, rapid reaction, high sensitivity, good specificity, etc.

Description

technical field [0001] The invention relates to the technical field of fluorescence immunochromatography in medical immunology, in particular to an NT-proBNP detection kit and a detection method capable of rapidly and accurately performing quantitative analysis on NT-proBNP. Background technique [0002] Heart failure is a group of syndromes caused by various structural or functional diseases of the heart that lead to impaired ventricular filling and / or ejection ability, and it is one of the most common causes of hospitalization or death in the elderly. With the aging of the population and the increase in the survival rate of myocardial infarction, heart failure is the only cardiovascular disease whose incidence and prevalence are still rising, and its prevention and treatment has become a hot spot in clinical cardiology research in recent years. Studies in recent years have shown that N-terminal pro-B-type natriuretic peptide (NT-proBNP) has a higher value in early diagnosi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/558G01N33/6893G01N2800/325G01N2800/50
Inventor 滕志涛于鸿翔陈萍萍
Owner WEIHAI NEOPROBIO
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