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Methoxynorepinephrine Luminescence Immunoassay Kit

A technology of norepinephrine and detection kit, applied in chemiluminescence/bioluminescence, measurement device, analysis by chemical reaction of materials, etc., can solve HPLC time-consuming, high cost, enzyme-linked immunoassay method Lack of accuracy and reproducibility issues to achieve high sensitivity and precision

Active Publication Date: 2021-05-25
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the mainstream detection methods of methoxynorepinephrine in the market mainly include high performance liquid chromatography (HPLC), enzyme-linked immunosorbent assay (ELISA), radioimmunoassay, but HPLC is time-consuming, costly and difficult to implement Batch assays where ELISA lacks accuracy and reproducibility

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  • Methoxynorepinephrine Luminescence Immunoassay Kit
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  • Methoxynorepinephrine Luminescence Immunoassay Kit

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Example 1 Preparation of methoxynorepinephrine luminescent immunoassay kit

[0032] 1. Preparation of solid phase carrier material

[0033] Preparation of magnetic microsphere suspension: first, wash the selected magnetic particle stock solution with 10 times the stock solution volume of PBS buffer for 2 to 5 times, then use EDC, NHS or glutaraldehyde to activate, and the activated magnetic microspheres are mixed with Antibodies with a concentration of 5-40 μg / mL are coated by any method of chemical linkage. After the coated magnetic microspheres are washed and sealed with a sealing solution, they are fixed to volume and packaged, and stored at 2-8 °C. spare.

[0034] This method can be used to prepare a, magnetic microspheres linked to the second antibody, b, magnetic microspheres linked to the anti-FITC antibody, c, magnetic microspheres linked to the magnetic microspheres suspension of the anti-methoxynorepinephrine antibody.

[0035] 2. Preparation of avidin-linke...

Embodiment 2

[0048] Embodiment 2 The using method of kit of the present invention

[0049] 1. Sample pretreatment: Take 10-50 μl of urine test sample into a 6 mL glass bottle, add 100-300 μl of acidification solution, acidify in a water bath (60-100°C) for 0.5-2 h, cool to room temperature, add 20- 100 μl of acylating agent was shaken on a shaker for 15-30 min, transferred to a cuvette, and detected using an AutoLumo automatic detection analyzer.

[0050] 2. Detection: Take the kit composed of avidin-linked tracer, methoxynorepinephrine antibody solution, common conventional substrate, and cleaning solution as an example: add the processed calibrator and sample into the cuvette respectively , the sample volume was 50 μl / well. Add 20 μl of magnetic particle suspension, 50 μl of sample, and 50 μl of antibody solution to each well, mix well and incubate at 37°C for 15 minutes, and wash with washing solution 6 times. Add 100 μl of avidin-linked tracer to each well, mix well and incubate at 3...

Embodiment 3

[0051] The performance evaluation of embodiment 3 kit of the present invention

[0052] 1. Sensitivity detection

[0053] Limit of Blank (LOB): 5 blank clinical samples with a value close to 0, each sample was repeated 3 times for a total of 4 days, and 60 data with non-negative results were obtained;

[0054] Line of Detection (LOD): After the LOB is determined, collect 5 clinical samples with a low value of 1 to 4 times the LOB, repeat 3 times for each sample, and do a total of 4 days to obtain 60 data;

[0055] Functional Sensitivity (FS): Using the data in the LOD experiment, 5 concentration samples were tested 3 times a day for a total of 4 days, and each sample obtained 12 results, and the mean, SD and CV% of each sample were calculated, and the nearest 20 The concentration of % is the functional sensitivity. The specific data are shown in Table 1.

[0056] Table 1 Sensitivity detection of the kit of the present invention

[0057]

[0058] It can be seen from the ...

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Abstract

The invention discloses a methoxynorepinephrine luminescent immunoassay kit, which includes a detection system and a sample pretreatment system; wherein the detection system includes a solid phase coated with an anti-methoxynorepinephrine antibody directly or indirectly Carrier, methoxynorepinephrine antibody solution, avidin-linked tracer solution and calibrator; sample pretreatment system includes acidifying solution, acylating agent and alkaline buffer. The present invention overcomes the shortcomings and deficiencies of the existing methoxynorepinephrine detection methods. Firstly, the methoxynorepinephrine in urine or blood plasma undergoes acidification and acylation pretreatment, and acylation with biotin at one end is adopted. Reagent acylation scheme, after the acylated methoxynorepinephrine is recognized and reacted by the antibody, it is combined with the method of avidin linked to the tracer label to accurately determine the methoxynorepinephrine in the sample content. The prepared kit has high sensitivity and precision, which can realize the automation of detection and facilitate the clinical diagnosis of pheochromocytoma.

Description

technical field [0001] The invention relates to biological detection technology, in particular to a methoxynorepinephrine luminescence immunoassay kit. Background technique [0002] Normetanephrine (NMN) is a metabolite of norepinephrine, and it is a useful indicator for the diagnosis of pheochromocytoma, the study of the etiology of hypertension and the study of sympathetic nerve function. NMN is a monoamine neurotransmitter belonging to catecholamines (CAs). It is an endogenous substance with strong physiological activity and plays an important role in brain and nerve signal transduction. NMN is an intermediate metabolite of adrenaline metabolism, which is only metabolized in the adrenal medulla and pheochromocytoma and paraganglioma (PPGL) tumors and persists at high concentrations, so it is a specific marker of PPGL. [0003] Among patients with adrenal tumors, 0.5% to 1% are patients with pheochromocytoma, and the pheochromocytoma cells continuously or intermittently r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/531G01N21/76
CPCG01N21/76G01N33/531G01N33/57488
Inventor 庄路阳马雷陈小玲陈飞杨敏乔晓芳李晓霞付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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