Enzymatic method for determining potassium ion content and potassium ion diagnosis kit

A diagnostic kit and potassium ion technology are applied in the field of medical examination and determination, which can solve the problems of complicated operation, insufficient time, and the inability of a biochemical analyzer to simultaneously measure the content of potassium ions and sodium ions, and achieve the effects of high specificity and accurate test results.

Inactive Publication Date: 2006-06-14
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because there are holes in the macrocyclic structure, the oxygen atoms in the molecule have unshared electron pairs that can be combined with metal ions. According to the size of the holes, metal ions of different diameters can be selectively combined to achieve the purpose of measuring the ion concentration (determination Blood potassium, generally using common crown ether anionic dyes for colorimetric quantification); atomic spectrophotometry can also be used to detect K in serum + 、Na + , but the operation is complicated, the error is large, and it is not as simple as the flame photometry
[0007] The method for measuring potassium ion content by enzymatic method has good consistency with flame photometer method or ion selective electrode method. This method has strong anti-interference and good stability, but it cannot simultaneously measure potassium ion and The respective content of sodium ions makes this method unable to be popularized and applied.

Method used

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  • Enzymatic method for determining potassium ion content and potassium ion diagnosis kit
  • Enzymatic method for determining potassium ion content and potassium ion diagnosis kit
  • Enzymatic method for determining potassium ion content and potassium ion diagnosis kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment one (single agent)

[0063] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage:

[0064] Glycine buffer 80mmol / l,

[0065] Adenosine diphosphate 3mmol / l,

[0066] Phosphoenolpyruvate 2mmol / l,

[0067] Glucose 5mmol / l,

[0068] thio-NADH 1mmol / l,

[0069] Pyruvate kinase 6000U / l,

[0070] Hexokinase 10000U / l,

[0071] Glucose phosphate dehydrogenase 10000U / l,

[0072] 6-Phosphogluconolactonase 10000U / l,

[0073] Phosphogluconate dehydrogenase 10000U / l,

[0074] Ethylene glycol 50% (accounting for the total volume of the reagent).

[0075] Set on the fully automatic biochemical analyzer (Hitachi-7080): temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test secondary wavelength 405nm or more, the volume ratio of the measured potassium ion sample to the reagent is 1:25, The reaction direction is positive reaction (absorbance rises, the same below).

[0076] Add the sample and the prepared si...

Embodiment 2

[0077] Embodiment two (two doses)

[0078] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage:

[0079] Reagent I——

[0080] Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0081] Adenosine diphosphate 5mmol / l,

[0082] Phosphoenolpyruvate 3mmol / l,

[0083] Glucose 13mmol / l,

[0084] NADPH 2mmol / l,

[0085] Hexokinase 15000U / l,

[0086] Glucose phosphate dehydrogenase 15000U / l,

[0087] 6-phosphogluconolactonase 20000U / l,

[0088] Phosphogluconate dehydrogenase 20000U / l,

[0089] Glycerol 50% (accounting for the total volume of reagent I);

[0090] Reagent II——

[0091]Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0092] Pyruvate kinase 9000U / l,

[0093] Ethylene glycol 50% (accounting for the total volume of reagent II).

[0094] Set on the automatic biochemical analyzer (Hitachi-7080): temperature 30°C, reaction time 15 minutes, test main wavelength 340nm, test secondary wavelength 405nm or more, the ratio between th...

Embodiment 3

[0096] Embodiment three (three doses)

[0097] Prepare the potassium ion diagnostic kit according to the following ingredients and dosage:

[0098] Reagent I——

[0099] Triethanolamine buffer 120mmol / l,

[0100] Adenosine diphosphate 7mmol / l,

[0101] Phosphoenolpyruvate 4mmol / l,

[0102] Glucose 20mmol / l,

[0103] thio-NADPH 3mmol / l,

[0104] Propylene glycol 20mmol / l;

[0105] Reagent II——

[0106] Triethanolamine buffer 120mmol / l,

[0107] Hexokinase 20000U / l,

[0108] Glucose phosphate dehydrogenase 20000U / l,

[0109] 6-phosphogluconolactonase 30000U / l,

[0110] Phosphogluconate dehydrogenase 30000U / l,

[0111] Propylene glycol 50% (accounting for the total volume of reagent II);

[0112] Reagent III——

[0113] Triethanolamine buffer 120mmol / l,

[0114] Pyruvate kinase 12000U / l,

[0115] Ethylene glycol 50% (accounting for the total volume of reagent III).

[0116] Set on the automatic biochemical analyzer (Hitachi-7080): temperature 25°C, reaction time 20 ...

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Abstract

The invention relates to enzyme method measuring kalium ion content method and its diagnosis kit. It is utilized kalium ion activation pyruvate kinase specificity to couple hexokinase, glucose phosphate dehydrogenase, 6-phosphogluconolactone and phosphogluconate dehydrogenase to react, reduce dichotomy type oxidizing coenzyme to dichotomy reducing type. To measure dominant wavelength 340nm absorbance rising speed can quantificationally reflect sample kalium ion content. The method has high specificity, good accuracy. The diagnosis kit can be made into bi-agent or tri-agent to reduce cross influence. The method can be quickly measured by ultraviolet/visible light analyzer or semi/full automatic biochemical analysis. Its measuring cost is low; and it is convenient for generalization and application.

Description

technical field [0001] The invention relates to a method for measuring potassium ion content in samples such as serum and plasma by enzymatic method, and a potassium ion diagnostic kit prepared by applying the method, which belongs to the technical field of medical inspection and determination. Background technique [0002] The content of potassium ions in serum (referred to as "serum potassium") has very important clinical significance. Under normal circumstances, there is a reasonable reference value range for blood potassium in the human body. Hypokalemia occurs when serum potassium falls below the lower reference limit due to renal tubular disease, hyperaldosteronism, hyperinsulinemia, metabolic alkalosis, or the use of insulin for diabetic ketosis, use of diuretics Potassium ions are transferred to the inside of cells during the treatment of certain diseases; when blood potassium is higher than the upper limit of the reference value, it may be caused by glomerular disea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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