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Determination of magnesium ion content from enzyme method and magnesium ion diagnostic reagent kit

A diagnostic kit and magnesium ion technology are applied in the field of medical testing and determination, which can solve problems such as differences in the accuracy of sensitivity test results, influence use and promotion, etc., and achieve the effects of low test cost, credible test results, and stable reagents.

Inactive Publication Date: 2006-05-31
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with different detection reagents and testing routes, the sensitivity of detection and the accuracy of detection results will be very different, which directly affects the use and promotion of this method. This is why people continue to research, develop and innovate in this field. the reason for

Method used

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  • Determination of magnesium ion content from enzyme method and magnesium ion diagnostic reagent kit
  • Determination of magnesium ion content from enzyme method and magnesium ion diagnostic reagent kit
  • Determination of magnesium ion content from enzyme method and magnesium ion diagnostic reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment one (single agent)

[0049] Prepare the serum magnesium ion diagnostic kit according to the following components and dosage:

[0050] Glycine buffer 80mmol / l,

[0051] Glycerol 10mmol / l,

[0052] Adenosine triphosphate 2mmol / l,

[0053] thio-NADH 0.2mmol / l,

[0054] Glycerol kinase 10000U / l,

[0055] Glycerol-3-phosphate dehydrogenase 10000U / l,

[0056] Ethylene glycol 50% (accounting for the total volume of the reagent).

[0057] Set on the 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 tested sample to the reagent is 1:25, and the reaction direction Negative reaction (decrease in absorbance, the same below).

[0058] Add the serum sample and the prepared single dose, and the two are automatically mixed in the analyzer to detect and record the decrease of the absorbance at 340nm. A total of 31 reading points were de...

Embodiment 2

[0059] Embodiment two (two doses)

[0060] Prepare the serum magnesium ion diagnostic kit according to the following components and dosage:

[0061] Reagent I——

[0062] Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0063] Glycerol 15mmol / l,

[0064] Adenosine triphosphate 5mmol / l,

[0065] NADPH 0.25mmol / l,

[0066] Glycerol-3-phosphate dehydrogenase 20000U / l,

[0067] Ethylene glycol 50% (accounting for reagent I total volume);

[0068] Reagent II——

[0069] Imidazole ~ hydrochloric acid buffer 100mmol / l,

[0070] Glycerol kinase 20000U / l,

[0071] Ethylene glycol 50% (accounting for the total volume of reagent II).

[0072] 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 of the total volume of the tested sample to reagent I and reagent II is 1:25, the ratio of the amount of reagent I to reagent II is 4:1, and the reaction directi...

Embodiment 3

[0074] Embodiment three (three doses)

[0075] Prepare the serum magnesium ion diagnostic kit according to the following components and dosage:

[0076] Reagent I——

[0077] Triethanolamine buffer 120mmol / l,

[0078] Glycerol 20mmol / l,

[0079] Adenosine triphosphate 8mmol / l,

[0080] thio-NADPH 0.3mmol / l,

[0081] Propylene glycol 20mmol / l;

[0082] Reagent II——

[0083] Triethanolamine buffer 120mmol / l,

[0084] Glycerol-3-phosphate dehydrogenase 30000U / l,

[0085] Propylene glycol 50% (accounting for the total volume of reagent II);

[0086] Reagent III——

[0087] Triethanolamine buffer 120mmol / l,

[0088] Glycerol kinase 30000U / l,

[0089] Ethylene glycol 50% (accounting for the total volume of reagent III).

[0090] Set on the automatic biochemical analyzer (Hitachi-7080): temperature 25°C, reaction time 20 minutes, test main wavelength 340nm, test secondary wavelength 405nm or more, the total volume of the tested sample and reagent I, reagent II and reagent I...

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Abstract

The invention is about the enzyme method of measuring magnesium ion and its diagnosis reagent box. Making use of the peculiarity that magnesium ion in the sample of plasma or serum and so on can activate the activation of glycerokinase, and producing glycerol-3-phosphoryl by reacting glycerol under the existence of adenosine triphosphate, and then causing coupled reaction with glycerol-3-phosphoryl dehydrogenase and transferring oxidized coenzyme to reduced coenzyme. Testing the descending range of dominant wave-length340nm absorbance and finally measuring the content of magnesium ion in the sample. This method has high specificity and would not be contaminated by material of internal and exogenous sources, and the result is precise and accurate. Diving the diagnosis reagent box into double-dose or three-dose can reduces the cross interaction of each element, keeps the stability of the reagent and deposits chronically. Using this method can realize the fast testing in common ultraviolet / visible light analyzer or semiautomatic / automatic analyzer and doesní»t require special or additional apparatus, so the cost is low. Thus, this method can be easily promoted and applied in the whole industry.

Description

technical field [0001] The invention relates to a method for enzymatically measuring magnesium ion content in samples such as plasma and serum, and a magnesium ion diagnostic kit prepared by applying the method, which belongs to the technical field of medical inspection and determination. Background technique [0002] At present, there are many methods for the determination of magnesium ion content in samples such as plasma and serum (referred to as magnesium determination), which can be summarized as follows: colorimetric method, fluorescence method, ion chromatography, ion selective electrode method (ISE), enzymatic method, Atomic absorption spectrophotometry (AAS), isotope dilution mass spectrometry (ID-MS), etc. Among them, the decisive method is ID-MS, followed by AAS. Although the results of these two methods are accurate, the equipment is complex and expensive, and they are not suitable for routine laboratory and automatic analysis. [0003] Colorimetry is the most w...

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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