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Epinephrine luminescence immunodetection kit

A detection kit, epinephrine technology, applied in biological testing, chemiluminescence/bioluminescence, measurement devices, etc., can solve the problems of time-consuming HPLC, lack of accuracy and reproducibility of enzyme-linked immunoassay, and high cost, Achieve high sensitivity and precision

Active Publication Date: 2019-03-08
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 luminescence immunodetection kit
  • Epinephrine luminescence immunodetection kit
  • Epinephrine luminescence immunodetection 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, 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 linking. After the coated magnetic microspheres are washed and sealed with a sealing solution, they are fixed to volume and subpackaged, 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-EITC antibody, c, magnetic particle suspension of magnetic particle-linked anti-adrenaline antibody.

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

[0036] First,...

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, and place on the shaker Shake for 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 10-10 min Transfer 100 μl to a 96 reaction cup, add 10-100 μl of methylase solution, shake on the oscillator 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, comm...

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] It can be seen from Table 1 that the concent...

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Abstract

The invention discloses an epinephrine luminescence immunodetection kit. The epinephrine luminescence immunodetection kit comprises a detection system and a sample pretreatment system, wherein the detection system comprises a solid phase carrier which is coated with an epinephrine antibody directly or indirectly, an epinephrine antibody solution, an avidin connection tracer solution and a calibration product; the sample pretreatment system comprises an enrichment material, an acylating agent and methylase. By adopting the epinephrine luminescence immunodetection kit, the defects and disadvantages in the existing epinephrine detection method are overcome. Firstly, epinephrine in urea or plasma is subjected to enrichment, acylation and methylation pretreatment, and is adsorbed and separatedwith a cis-diol specific affine medium. The scheme of acylation with the acylating agent having biotin at one end is adopted, so that the content of epinephrine in a sample is accurately determined inconjunction with avidin connected with a tracing marker after the acylated or methylated epinephrine is recognized by an antibody and reacted. The kit has the advantages of high sensitivity and precision, detection automation is realized by aluminescence technology, and the clinical diagnosis of pheochromocytoma is assisted.

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 Applications(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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