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Pheochromocytoma diagnostic kit and using method thereof

A technology for pheochromocytoma and diagnostic kits, applied in the field of providing a kit, which can solve the problems of high false positive rate and achieve the effect of simple reagents, low price and high diagnostic efficiency

Active Publication Date: 2017-12-12
重庆医科大学国际体外诊断研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, pheochromocytoma is mainly diagnosed clinically by detecting the levels of free NMN and MN in plasma, but the false positive rate is high

Method used

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  • Pheochromocytoma diagnostic kit and using method thereof
  • Pheochromocytoma diagnostic kit and using method thereof
  • Pheochromocytoma diagnostic kit and using method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: the preparation of pheochromocytoma diagnostic kit

[0045] 1. Instruments and equipment

[0046] 1. Analytical balance: Sensitivity is 0.0001g

[0047] 2. Microporous membrane: pore size 0.22μm

[0048] 3. Agilent 1100 High Performance Liquid Chromatograph

[0049] 4. Millipore ultrapure water filtration device and Millipore vacuum pump (Millipore Corporation, USA)

[0050] 5. HIMAC-cf 15 low temperature desktop centrifuge

[0051] 2. Reagents and solutions: the components of the kit are shown in Table 2

[0052] Table 2 Kit components

[0053]

[0054] The preparation method of the above-mentioned kit specifically includes:

[0055] (1) Sample pretreatment reagents:

[0056] ①Prepare 10% ammonia water methanol solution (volume ratio 3:1); filter with 0.22 μm organic membrane, and store in 4°C refrigerator for later use.

[0057] ② Prepare 0.18mol / L potassium hydroxide methanol solution; filter with 0.22μm organic membrane, and store in 4°C ref...

Embodiment 2

[0065] Example 2: Use of Pheochromocytoma Diagnostic Kit

[0066] 1. Urine Sample Pretreatment

[0067] Take 500μL random urine, add 5mL ultrapure water, 200μL 0.2mol / L acetic acid, and mix well. Use 5mL of 10% ammonia water-methanol (volume ratio 1:3), 2mL of 0.18mol / L potassium hydroxide methanol solution, and 2mL of ultrapure water to activate the solid-phase extraction column in turn, mix the above samples and load them, and control the flow rate to 1 -2mL / min. Rinse the cartridge sequentially with 4mL 10mmol / L acetic acid-methanol (volume ratio 9:1), 4mL 10mmol / L ammonium phosphate, and 4mL ultrapure water. 2mL ammonia water-methanol solution was used for elution, the eluate was spin-dried in vacuum and then redissolved with 200μL 0.20mol / L acetic acid solution.

[0068] 2. Liquid chromatography conditions

[0069] Mobile phase: mobile phase A is 70mmol / L NaH 2 PO 4 solution, the mobile phase B is methanol (chromatographically pure), and a gradient elution method is...

Embodiment 3

[0078] This example is to investigate the precision of the kit of the present invention for determining NMN, MN and 3-MT in random urine. Real samples were used to investigate the precision, that is, the urine of patients with pheochromocytoma and the urine of normal people were selected, and the measurement was repeated 5 times a day for 5 consecutive days, and the ratio of the analyte to creatinine was calculated to evaluate the precision. See Table 4. The coefficients of variation are all equal to or less than 5.6%, indicating that the present invention has good precision.

[0079] Table 4 Precision (n=5)

[0080]

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Abstract

The invention belongs to the technical field of detection and in particular relates to a pheochromocytoma diagnostic kit and a using method thereof. According to the technical scheme in the invention, normetanephrine (NMN), metanephrine (MN), 3-methyltyramine (3-MT) and creatinine (Cr) in random urine can be detected by adopting high performance liquid chromatography, and pheochromocytoma is diagnosed by utilizing a ratio of the previous three to-be-detected substances to a peak area of the creatinine. The kit disclosed by the invention comprises the following reagents: a sample pretreatment reagent, a standard stock solution and a mobile phase. The pheochromocytoma is diagnosed by directly utilizing the ratio of the three to-be-detected substances to the peak area of the creatinine, and the pheochromocytoma diagnostic kit is simple in reagents, low in price, convenient to use and high in cost performance.

Description

technical field [0001] The invention provides a kit, which mainly relates to the diagnosis of pheochromocytoma and belongs to the technical field of detection. Background technique [0002] Pheochromocytoma is an endocrine tumor originating from the adrenal medulla and the extra-adrenal sympathetic nervous system. It is characterized by the secretion of a large amount of catecholamines, and typically manifests as the triad of "palpitations, sweating, and headache" and secondary hypertension. The greatest harm of the disease is concurrent hypertensive crisis, acute left heart failure and cerebral hemorrhage, and severe cases are life-threatening. Pheochromocytoma is closely related to the metabolism of catecholamines. Catecholamine is a neurological substance containing catechol and amine groups, including norepinephrine (Norepinephrine, NE), epinephrine (Epinephrine, E) and dopamine (Dopamine, DA). Under the catalysis of Catechol-O-methyltransferase (COMT), DA, NE and E ar...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/74
CPCG01N30/02G01N30/74
Inventor 丁敏邹文毕甄乾娜张晓清吕文静陈霄韩润川
Owner 重庆医科大学国际体外诊断研究院
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