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Luminescence immunoassay kit for metanephrine

A metanephrine, detection kit technology, applied in chemiluminescence/bioluminescence, measurement device, analysis by chemical reaction of materials, etc., can solve the problem of time-consuming HPLC and the lack of accuracy of enzyme-linked immunoassay To solve the problems of high reliability and reproducibility, and high cost, to achieve the effect of 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 methoxyepinephrine 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 ELISA lacks accuracy and reproducibility

Method used

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  • Luminescence immunoassay kit for metanephrine
  • Luminescence immunoassay kit for metanephrine
  • Luminescence immunoassay kit for metanephrine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Preparation of methoxyepinephrine luminescent 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, then activate it with EDC, NHS or glutaraldehyde, and the activated magnetic microspheres Coat the antibody with a concentration of 5-40 μg / mL by any method of chemical connection. 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 particle-linked second antibody, b, magnetic particle-linked anti-EITC antibody, c, magnetic particle suspension of magnetic particle-linked anti-methoxyadrenaline antibody.

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

[0036] First, according to the formul...

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 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 consisting of avidin-HRP solution, methoxyepinephrine antibody solution, common conventional substrate, and cleaning solution as an example: add the processed calibrator and sample into the cuvette respectively, add 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 enzyme conjugate to each well, mix well and incubate at 37 °...

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; the specific data are shown in Table 1.

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

[0057]

[0058] The results in Table 1 sh...

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Abstract

The invention discloses a luminescence immunoassay kit for metanephrine. The luminescence immunoassay kit comprises a detection system and a sample pretreatment system, wherein the detection system comprises a solid phase carrier which is directly or indirectly coated with a metanephrine antibody, a metanephrine antibody solution, an avidin link tracer solution and a calibration product; the sample pretreatment system comprises acidizing fluid, an acylating agent and alkaline buffer. According to the luminescence immunoassay kit disclosed by the invention, the defects and deficiency of an existing detection method of the metanephrine are overcome; the metanephrine in urine or plasma is subjected to acidifying and acylating pretreatment; by adopting an acylating scheme of the acylating agent of which one end is biotin, the acylated metanephrine is identified by an antibody and reacts, and then the content of the metanephrine in a sample is accurately determined through combination witha method of a tracer linked avidin. The prepared kit has high sensitivity and precision; a luminous technology is utilized to realize detection automation and assists in clinical diagnosis of pheochromocytoma.

Description

technical field [0001] The invention relates to biological detection technology, in particular to a methoxyepinephrine luminescence immunoassay kit. Background technique [0002] Methoxyepinephrine (metanephrine, MN) is a metabolite of epinephrine, which is a useful indicator for the diagnosis of pheochromocytoma, the study of the etiology of hypertension and the study of sympathetic nerve function. MN 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. MN is an intermediate metabolite of adrenaline metabolism, which is only metabolized in the adrenal medulla, pheochromocytoma and paraganglioma (PPGL) 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 rele...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/531G01N33/543G01N21/76
CPCG01N21/76G01N33/531G01N33/54326G01N33/57488
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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