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Epinephrine Luminescent Immunoassay Kit

A detection kit, epinephrine technology, applied in biological testing, chemiluminescence/bioluminescence, measurement devices, etc., can solve the problems of time-consuming HPLC, high cost, and lack of accuracy and reproducibility of enzyme-linked immunoassays , 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 epinephrine 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 achieve batch determination, while enzyme Linked immunoassays lack accuracy and reproducibility

Method used

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  • Epinephrine Luminescent Immunoassay Kit
  • Epinephrine Luminescent Immunoassay Kit
  • Epinephrine Luminescent Immunoassay Kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Preparation of epinephrine luminescence immunoassay kit

[0032] 1. Preparation of solid phase carrier material

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

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

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

[0036] First, according t...

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, epinephrine antibody solution, common conventional substrat...

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, closest to 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] It can be seen from Table 1 that the concentratio...

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Abstract

The invention discloses an epinephrine 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-adrenaline antibody, an adrenaline antibody solution, and avidin Connect tracer solution and calibrator; sample pretreatment system includes enrichment material, acylating agent, methylase. The present invention overcomes the shortcomings and deficiencies of the existing adrenaline detection methods. Firstly, the adrenaline in urine or plasma is enriched, acylated and methylated before treatment, and the adrenaline is adsorbed and separated by using a cis-diol-specific affinity medium. , adopt the acylation scheme of acylating agent with biotin at one end, make the acylated and methylated adrenaline recognized and reacted by the antibody, and combine with the method of avidin linked to the tracer label to accurately measure the avidin in the sample of adrenaline content. 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 an epinephrine luminescence immunoassay kit. Background technique [0002] Adrenaline (adrenaline) is a hormone secreted by the human adrenal gland, and it is a key neurotransmitter in the hypothalamus and pituitary gland. Further, through the action of phenylethylamine N-methyl transferase (phenylethanolamine N-methyl transferase, PNMT), norepinephrine is methylated to form epinephrine. Epinephrine has a similar chemical structure to dopamine and norepinephrine, which are compounds composed of a benzene ring (catechol nucleus) containing two adjacent hydroxyl groups and a side chain containing an amino group. [0003] Among patients with adrenal tumors, 0.5% to 1% are patients with pheochromocytoma, and the pheochromocytoma cells continuously or intermittently release a large amount of catecholamines, so the main clinical symptoms are excessive catecholamines, causing persiste...

Claims

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Application Information

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