Norepinephrine Luminescent Immunoassay Kit

A norepinephrine and detection kit technology, which is applied in biological testing, chemiluminescence/bioluminescence, measuring devices, etc. It can solve the problems of time-consuming HPLC, lack of accuracy and reproducibility, and high cost of enzyme-linked immunoassay. problem, to achieve the effect of high sensitivity and precision

Active Publication Date: 2021-05-28
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 norepinephrine in the market mainly include high performance liquid chromatography (HPLC), enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay, but HPLC is time-consuming, costly and difficult to achieve batch determination. Enzyme-linked immunoassays lack accuracy and reproducibility

Method used

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  • Norepinephrine Luminescent Immunoassay Kit
  • Norepinephrine Luminescent Immunoassay Kit
  • Norepinephrine Luminescent Immunoassay Kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Preparation of norepinephrine luminescent immunoassay kit

[0032] 1. Preparation of solid phase carrier material

[0033] Preparation of magnetic microsphere suspension: firstly, the selected magnetic microsphere stock solution was washed with 10 times the stock solution volume of PBS buffer for 2 to 5 times, and then activated with EDC, NHS or glutaraldehyde, and the activated magnetic microspheres were 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 for later use. .

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

[0035] 2. Preparation of avidin-linked tracer solut...

Embodiment 2

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

[0049] 1. Sample pretreatment: Take 10-50 μl of urine test sample in cis-diol-specific affinity medium, add 1000-5000 μl of extraction buffer, shake on the shaker for 0.5-2 h, remove the liquid and pat dry , add 2 mL of purified water, shake on the shaker for 2-10 min, remove the liquid, then add 100-500 μl of extraction buffer, add 10-100 μl of acylation reagent to each well, mix immediately, shake on the shaker 10-50 min, remove the liquid and pat dry, add 2 mL of purified water, shake on the shaker for 2-10 min, absorb 100-600 μl release buffer, shake on the shaker for 0.5-2 h, take the supernatant for 10-100 μl Transfer μl to a 96 reaction cup, add 10-100 μl of methylase solution, shake on a shaker for 0.5-2 h, and use the AutoLumo automatic detection analyzer for detection.

[0050] 2. Detection: Take the kit composed of avidin-linked tracer, norepinephrine antibody solution, common conventional subst...

Embodiment 3

[0051] Embodiment 3 Performance evaluation of the 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; as shown in Table 1.

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

[0057]

[0058] Table 1 shows that the concentration that ca...

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Abstract

The invention discloses a norepinephrine luminescent immunoassay kit, which includes a detection system and a sample pretreatment system; wherein the detection system includes a solid-phase carrier directly or indirectly coated with an anti-norepinephrine antibody, norepinephrine Antibody solution, avidin-linked tracer solution and calibrator; sample pretreatment system includes enrichment material, acylating agent, methylase. The present invention first enriches, acylates, and pre-methylates norepinephrine in urine or plasma, uses cis-diol-specific affinity media to adsorb and separate norepinephrine, and adopts an acylation method in which one end is biotin The reagent acylation scheme, after the acylated and methylated norepinephrine is recognized and reacted by the antibody, is combined with the method of avidin linked to the tracer label to accurately determine the norepinephrine content in the sample. The kit of the invention has high sensitivity and precision, uses luminescence technology to realize automatic detection, and assists clinical diagnosis of pheochromocytoma.

Description

technical field [0001] The invention relates to biological detection technology, in particular to a norepinephrine luminescent immunoassay kit. Background technique [0002] Norepinephrine, scientific name 1-(3,4-dihydroxyphenyl)-2-aminoethanol, is a substance formed by removing the N-methyl group of epinephrine, and also belongs to catecholamine in chemical structure. It is not only a neurotransmitter, which is mainly synthesized and secreted by sympathetic postganglionic neurons and adrenergic nerve endings in the brain, and is the main transmitter released by the latter, but also a hormone, which is synthesized and secreted by the adrenal medulla, but The content is less. Circulating norepinephrine comes primarily from the adrenal medulla. [0003] Certain tumors can lead to abnormal metabolism of catecholamines, such as pheochromocytoma and paraganglioma. Pheochromoma cells continuously or intermittently release a large amount of catecholamines, so the main clinical s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531G01N33/543G01N33/74G01N21/76
CPCG01N21/76G01N33/531G01N33/54326G01N33/74
Inventor 庄路阳马雷陈小玲陈飞肖静乔晓芳李晓霞付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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