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 immuno

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

Example Embodiment

[0031] Example 1 Preparation of methoxy adrenaline luminescence immunoassay kit

[0032] 1. Preparation of solid phase carrier material

[0033] Preparation of magnetic microsphere suspension: first wash the selected magnetic microsphere stock solution with 10 times the stock solution volume of PBS buffer for 2 to 5 times, then use EDC, NHS or glutaraldehyde for activation. The activated magnetic microsphere Coated with antibodies with a concentration of 5-40 μg / mL by chemical linking or any method. The coated magnetic microspheres are washed and sealed with a sealing solution, then fixed to volume and aliquoted, and stored at 2-8 ℃ spare.

[0034] This method can be used to prepare a magnetic particle suspension with magnetic particles connected to a second antibody, b, magnetic particles connected to an anti-EITC antibody, and c, magnetic particles connected to an anti-methoxy adrenaline antibody.

[0035] 2. Prepare avidin-linked tracer solution

[0036] First follow the formula Tr...

Example Embodiment

[0048] Example 2 Method of using the kit of the present invention

[0049] 1. Sample pretreatment: Take 10-50 μl of urine test sample into 6 mL glass bottle, add 100-300 μl acidification solution, acidify in water bath (60-100℃) for 0.5-2 h, cool to room temperature, add 20- 100μl of acylating agent, shake on a shaker for 15-30 minutes, transfer to the reaction cup, and use AutoLumo automatic detection analyzer for detection.

[0050] 2. Detection: Take a kit consisting of avidin-HRP solution, methoxy adrenaline antibody solution, common conventional substrates, and cleaning solution as an example: add processed calibrators and samples to the reaction cup, and add The sample volume is 50μl / well. Add 20 μl of magnetic particle suspension, 50 μl of sample, and 50 μl of antibody solution to each well. After mixing, incubate at 37 ℃ for 15 minutes, and wash with lotion 6 times. Add 100 μl of enzyme conjugate to each well, incubate at 37 ℃ for 17 min after mixing, wash 6 times with lo...

Example Embodiment

[0051] Example 3 Performance evaluation of the kit of the present invention

[0052] 1. Sensitivity detection

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

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

[0055] Functional Sensitivity (FS): Using the data in the LOD experiment, 5 concentration samples are measured 3 times a day for a total of 4 days. Each sample gets 12 results. Calculate the mean, SD and CV% of each sample, which is the closest to 20 % Concentration is the functional sensitivity; the specific data is shown in Table 1.

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

[0057]

[0058] The results in Table 1 show that the concentration that can be accurately detected in the first batch is...

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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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