Magnesium ion detection reagent, kit and detection method

Abstract

The invention belongs to the technical field of biological detection, and relates to a magnesium ion detection reagent which comprises a reagent R1 and a reagent R2, the reagent R1 comprises a first buffer solution, glucose, ATP and NADP < + >; and the reagent R2 comprises the following components: a second buffer solution, glucokinase and glucose 6-phosphate dehydrogenase. The invention also provides a magnesium ion detection kit and a detection method. According to the present invention, the stability of the reagent can be improved so as to improve the detection accuracy of the reagent, the biological safety of the raw material of the detection reagent is good, and the magnesium ion detection operation process is simple, rapid and practical so as to be suitable for the clinical routine examination.

Description

Technical Field

[0001] The present application relates to the field of biological detection technology, and in particular to a magnesium ion detection reagent, a kit and a detection method. Background Art

[0002] Magnesium (Mg) is one of the most abundant cations in the body. The Mg content in an adult is 0.8-1.2 mol, of which 50% is in bones, 45% in intracellular fluid, and 5% in extracellular fluid. The liver, kidneys, and muscles contain more Mg. The content of Mg in intracellular fluid ranks second only to potassium, and its concentration is about 10 times that of extracellular fluid. In extracellular fluid, the content of Mg ranks fourth only to sodium, potassium, and calcium. In many physiological and chemical processes, Mg participates in reactions and plays an important role, such as being an activator of many enzymes. Mg is also an essential element for the composition of DNA, RNA, and ribosome macromolecules, and is also an important element for maintaining normal nerve function and muscles.

[0003] Traditionally, Mg is measured mainly for the diagnosis of kidney disease. In recent years, the clinical relationship between Mg deficiency and hypertension, myocardial infarction, diabetes and other diseases has become increasingly clear, attracting widespread attention. At present, the main clinical methods for Mg determination are atomic absorption spectrometry, xylene blue (XB) method and glucokinase (GlcK) method.

[0004] Among them, atomic absorption methods can perform high-precision measurements, but they require special expensive equipment and operating skills, have high detection costs, and cannot achieve fast and convenient detection. The XB method is the most widely used, easy to operate, low cost, and can be used in automatic biochemical analysis systems. However, since it is a photometric method in the visible light region, it is easily affected by other positive ions, and the reagents are unstable, so errors will occur due to the absorption of components of similar hues, and the detection accuracy is poor.

[0005] The GlcK method is an enzymatic method for detecting magnesium ions. Under the catalysis of GlcK (glucokinase), glucose and ATP-Mg complex undergo phosphorylation reaction to generate glucose-6-phosphate and ADP. Glucose-6-phosphate is dehydrogenated under the catalysis of 6-phosphoglucose dehydrogenase (G6PDH) to generate 6-phosphogluconic acid, while NADP + Reduced to NADPH. The principle of the enzymatic method is to use glucose kinase to react with the Mg-ATP complex. Its activity depends on the amount of the complex. The GlcK method has a higher specificity for Mg ions. However, under normal circumstances, NADPH is relatively stable only when the pH value of the aqueous solution is greater than 9. At present, the pH value of existing reagents is relatively low, resulting in the enzyme method The stability of magnesium ion reagents is not enough to meet the needs of clinical measurement samples. Summary of the invention

[0006] The purpose of the embodiments of the present application is to provide a magnesium ion detection reagent, a kit and a detection method to solve the technical problem of insufficient stability of reagents for enzymatic determination of magnesium ions in the prior art.

[0007] In order to solve the above technical problems, the present application embodiment provides a magnesium ion detection reagent, which adopts the following technical solution:

[0008] Including R1 reagent and R2 reagent;

[0009] The components of the R1 reagent include a first buffer, glucose, ATP and NADP + ; The components of the R2 reagent include a second buffer, glucose kinase and 6-phosphate glucose dehydrogenase.

[0010] Furthermore, the first buffer and the second buffer are independently selected from one of Tris-HCl buffer, HEPES buffer, PIPES buffer, CAPS buffer, glycine buffer, triethanolamine-HCl buffer, citric acid-trisodium citrate buffer, phosphate buffer and potassium hydrogen phthalate-hydrochloric acid buffer.

[0011] Furthermore, the components of the R1 reagent and the R2 reagent also independently include a surfactant, a stabilizer and a preservative.

[0012] Further, the surfactant is selected from at least one of emulsifiers OP, SDS, Tween20, Tween80, Brij35, Brij98, Tritonx-100, Tritonx-405, B66, A90 and 15-S9;

[0013] The stabilizer is selected from at least one of NaCl, sucrose, polyols, bovine serum albumin and crown ethers;

[0014] The preservative is selected from one of sodium azide, ProClin series, thimerosal, gentamicin and sodium benzoate.

[0015] Further, the concentrations of the first buffer and the second buffer are independently 90 to 110 mmol / L; and / or

[0016] The concentration of glucose is 9-10 mmol / L; and / or

[0017] The concentration of ATP is 9-10 mmol / L; and / or

[0018] The NADP + The concentration is 0.5-0.8 mmol / L; and / or

[0019] The concentration of glucokinase is 3-5 KU / L; and / or

[0020] The concentration of the 6-phosphate glucose dehydrogenase is 6-10 KU / L; and / or

[0021] The concentration of the surfactant is 4 to 6 g / L; and / or

[0022] The concentration of the stabilizer is 0.5 to 1.5 g / L; and / or

[0023] The concentration of the preservative is 0.5-1.0 g / L.

[0024] Furthermore, the pH value of the first buffer solution is 6.0-7.0, and the pH value of the second buffer solution is 8.0-9.0.

[0025] In order to solve the above technical problems, an embodiment of the present application also provides a magnesium ion detection kit, which includes the magnesium ion detection reagent as described above.

[0026] In order to solve the above technical problems, the present application also provides a method for detecting magnesium ions, which adopts the following technical solution:

[0027] Step S10, mixing the sample to be tested with the R1 reagent and the R2 reagent, wherein the components of the R1 reagent include a first buffer, glucose, ATP and NADP + ; The components of the R2 reagent include a second buffer, glucokinase and 6-phosphoglucokinase;

[0028] Step S20, the magnesium ions in the sample to be tested form an ATP-Mg complex with ATP;

[0029] Step S30, under the catalytic action of glucokinase, the ATP-Mg complex reacts with glucose to obtain glucose-6-phosphate and ADP-Mg, and the ADP-Mg immediately dissociates into magnesium ions and ADP, and the dissociated magnesium ions are circulated for step S20;

[0030] Step S40, glucose-6-phosphate and NADP + 6-phosphogluconate and NADPH are generated under the action of 6-phosphogluconate dehydrogenase;

[0031] Step S50, measuring NADPH, and determining the magnesium ion content in the sample to be tested according to the increase rate of NADPH.

[0032] Compared with the prior art, this application has the following beneficial effects:

[0033] The magnesium ion detection reagent provided in the present application includes R1 reagent and R2 reagent, and the components of R1 reagent include a first buffer, glucose, ATP and NADP + When the R1 reagent is added to the sample, ATP and magnesium ions are fully complexed; because the R1 reagent contains glucose, in order to avoid the glycosylation reaction between glucose kinase and 6-phosphate glucose dehydrogenase and glucose, which leads to a decrease in enzyme activity, these two enzymes are placed in the R2 reagent. The components of the R2 reagent include the second buffer, glucose kinase and 6-phosphate glucose dehydrogenase;

[0034] The present application stores the magnesium ion detection reagent as two different reagents, which can improve the stability of the reagents and thus improve the detection accuracy of the reagents. Secondly, the raw materials of the detection reagents have good biosafety. At the same time, the operation process of detecting magnesium ions is simple, fast, and practical, and is suitable for routine clinical examinations. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the scheme in the present application, a brief introduction is given below to the drawings required for use in the description of the embodiments of the present application. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0036] Figure 1 is a flow chart of an embodiment of a method for detecting magnesium ions according to the present application;

[0037] Figure 2 is a schematic diagram of the correlation results of Example 1 of the present application;

[0038] Figure 3 is a schematic diagram of the correlation results of Example 2 of the present application;

[0039] Figure 4 It is a schematic diagram of the correlation results of Example 3 of the present application. DETAILED DESCRIPTION

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by technicians in the technical field of the present application; the terms used in the specification of the application herein are only for the purpose of describing specific embodiments and are not intended to limit the present application; the terms "including" and "having" and any variations thereof in the specification and claims of the present application and the above-mentioned drawings are intended to cover non-exclusive inclusions. The terms "first", "second", etc. in the specification and claims of the present application or the above-mentioned drawings are used to distinguish different objects, not to describe a specific order.

[0041] Reference to "embodiments" herein means that a particular feature, structure, or characteristic described in conjunction with the embodiments may be included in at least one embodiment of the present application. The appearance of the phrase in various locations in the specification does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment that is mutually exclusive with other embodiments. It is explicitly and implicitly understood by those skilled in the art that the embodiments described herein may be combined with other embodiments.

[0042] In order to enable those skilled in the art to better understand the solution of the present application, the technical solution in the embodiments of the present application will be clearly and completely described below in conjunction with the accompanying drawings.

[0043] The present application provides a magnesium ion detection reagent, including R1 reagent and R2 reagent.

[0044] The components of R1 reagent include first buffer, glucose, ATP and NADP + ; The components of R2 reagent include a second buffer, glucose kinase (GlcK) and glucose-6-phosphate dehydrogenase (G6PDH).

[0045] ATP is adenosine triphosphate; NADP is + Nicotinamide adenine dinucleoside phosphate is the oxidized form of reduced coenzyme II (NADPH).

[0046] This application utilizes the characteristic of magnesium ions activating the activity of glucokinase, and the detection principle is as follows:

[0047]

[0048]

[0049] Magnesium ions first form an ATP-Mg complex with ATP. Under the catalytic action of GlcK, the ATP-Mg complex and glucose are converted into 6-phosphate glucose, and ADP-Mg is generated at the same time. The ADP-Mg produced in this reaction immediately dissociates into Mg and ADP, and Mg forms an ATP-Mg complex with ATP again. The 6-phosphate glucose produced by the GlcK-catalyzed reaction is in the presence of NADP + In the presence of magnesium, it is converted into 6-phosphogluconic acid through the catalysis of 6-phosphoglucose dehydrogenase (G6PDH), and NADPH is produced at the same time. NADPH has a very obvious absorption peak at a wavelength of 340nm. The absorbance change before and after the reaction is proportional to the content of magnesium ions. Therefore, by measuring the increase rate of absorbance of NADPH at a wavelength of 340nm, the content of magnesium ions can be indirectly measured.

[0050] At present, the XB method is commonly used for the detection of magnesium ions. The pH of this method needs to react under alkaline conditions (pH>10). The carbon dioxide in the air will be absorbed by the reagent, causing the pH to drop, which will eventually affect the stability of the reagent after opening the bottle. + It has better stability under acidic conditions (pH < 7), can eliminate the neutralization effect of carbon dioxide, and can meet the needs of clinical measurement samples.

[0051] In some embodiments, the first buffer and the second buffer are independently selected from one of Tris-HCl buffer, HEPES (4-hydroxyethylpiperazineethanesulfonic acid) buffer, PIPES (piperazine-1,4-diethanesulfonic acid) buffer, CAPS (3-(cyclohexylamine)-1-propanesulfonic acid) buffer, glycine buffer, triethanolamine-HCl buffer, citric acid-trisodium citrate buffer, phosphate buffer and potassium hydrogen phthalate-hydrochloric acid buffer.

[0052] In some embodiments, the pH value of the first buffer is 6.0-7.0, preferably 6.2, and the pH value of the second buffer is 8.0-9.0, preferably 8.5.

[0053] Buffers are used to maintain the stability of reagents and to provide a stable reaction environment.

[0054] In some embodiments, the components of the R1 agent and the R2 agent further independently include a surfactant, a stabilizer, and a preservative.

[0055] In this embodiment, the buffer, surfactant, stabilizer and preservative can improve the accuracy and stability of the detection reagent and eliminate various external interferences to the measurement results.

[0056] In some embodiments, the surfactant is selected from at least one of emulsifiers OP, SDS, Tween 20, Tween 80, Brij 35, Brij 98, Triton x-100, Triton x-405, B66, A90 and 15-S-9.

[0057] Among them, emulsifier OP is a liquid with high viscosity. It belongs to non-ionic surfactant, and its main chemical component is condensation product of alkylphenol and ethylene oxide; SDS is the abbreviation of Sodium dodecyl sulfate, which is sodium dodecyl sulfate; Brij35 represents dodecyl polyethylene glycol ether; Brij98 represents oleyl alcohol polyoxyethylene ether; Tritonx-100 represents polyethylene glycol octylphenyl ether; Tritonx-405 represents polyoxyethylene isooctylcyclohexyl ether; B66 represents polyoxyethylene tribenzyl phenyl ether; A90 represents polyoxyethylene distyrenated phenyl ether; 15-S-9 represents TERGITOL alkoxy polyethylene hydroxyethanol.

[0058] The stabilizer is selected from at least one of NaCl, sucrose, polyol, bovine serum albumin (BSA) and crown ether.

[0059] Among them, NaCl can increase the ion concentration and improve the stability of the ATP-Mg complex; sugars (such as sucrose) can protect the enzyme activity; polyols (such as glycerol), BSA, crown ethers, etc. can co-dissolve with proteins without reacting with them, thereby enhancing the stability of the protein and further protecting the enzyme activity.

[0060] The preservative is selected from sodium azide (NaN 3 ), one of the ProClin series, thimerosal, gentamicin, and sodium benzoate, used to inhibit the growth of microorganisms in the reagent.

[0061] Among them, the ProClin series preservatives include but are not limited to ProClin150, ProClin200, ProClin300, ProClin950 and ProClin5000.

[0062] In some embodiments, the concentrations of the first buffer and the second buffer are independently 90 to 110 mmol / L; and / or

[0063] The concentration of glucose is 9-10 mmol / L; and / or

[0064] ATP concentration is 9-10 mmol / L; and / or

[0065] NADP + The concentration is 0.5-0.8 mmol / L; and / or

[0066] The concentration of glucose dehydrogenase is 3-5 KU / L; and / or

[0067] The concentration of glucose-6-phosphate in glucose-6-phosphate dehydrogenase is 6 to 10 KU / L; and / or

[0068] The concentration of the surfactant is 4 to 6 g / L; and / or

[0069] The concentration of the stabilizer is 0.5 to 1.5 g / L; and / or

[0070] The concentration of the preservative is 0.5-1.0 g / L.

[0071] The concentration of the first buffer is preferably 100 mmol / L; the concentration of the second buffer is preferably 100 mmol / L; the concentration of glucose is preferably 10 mmol / L; the concentration of ATP is preferably 10 mmol / L; and the concentration of NADP is preferably 10 mmol / L. + The concentration of is preferably 0.7mmol / L; the concentration of glucose kinase is preferably 4KU / L; the concentration of 6-phosphoglucoside dehydrogenase is preferably 8KU / L; the concentration of surfactant is preferably 5g / L; the concentration of stabilizer is preferably 1g / L; the concentration of preservative is preferably 0.8g / L.

[0072] In some embodiments, the magnesium ion detection reagent is composed of R1 reagent and R2 reagent, wherein the components of R1 reagent are a first buffer, glucose, ATP, NADP + , surfactant, stabilizer and preservative; the components of R2 reagent are composed of a second buffer, glucose kinase, 6-phosphate glucose dehydrogenase, surfactant, stabilizer and preservative.

[0073] The magnesium ion detection reagent provided in the present application, including R1 reagent and R2 reagent, is a two-point endpoint determination reagent, which can improve the stability of the reagent and thus improve the detection accuracy of the reagent. Secondly, the raw materials of the detection reagent have good biosafety. At the same time, the operation process of detecting magnesium ions is simple, fast, and practical, and is suitable for routine clinical examinations.

[0074] Based on the above-mentioned magnesium ion detection reagent, the present application also provides a magnesium ion detection kit, which includes the magnesium ion detection reagent as described above, can improve the accuracy of magnesium ion detection, the detection process is simple and convenient, and the detection efficiency is high.

[0075] refer to Figure 1 , shows a flow chart of an embodiment of a method for detecting magnesium ions according to the present application, comprising the following steps:

[0076] Step S10, mixing the sample to be tested with the R1 reagent and the R2 reagent, wherein the components of the R1 reagent include a first buffer, glucose, ATP and NADP + ; The components of R2 reagent include a second buffer, glucokinase and 6-phosphoglucokinase;

[0077] Step S20, magnesium ions in the sample to be tested form ATP-Mg complex with ATP;

[0078] Step S30, under the catalytic action of glucokinase, the ATP-Mg complex reacts with glucose to obtain glucose-6-phosphate and ADP-Mg, wherein the ADP-Mg immediately dissociates into magnesium ions and ADP, and the dissociated magnesium ions are circulated for step S20;

[0079] Step S40, glucose-6-phosphate and NADP + 6-phosphogluconate and NADPH are generated under the action of 6-phosphogluconate dehydrogenase;

[0080] Step S50, measuring NADPH, and determining the magnesium ion content in the sample to be tested according to the increase rate of NADPH.

[0081] NADPH has a very obvious absorption peak at 340nm wavelength, and the change in absorbance before and after the reaction is proportional to the content of magnesium ions. Therefore, by measuring the increase rate of absorbance of NADPH at 340nm wavelength, the content of magnesium ions in the sample to be tested can be indirectly measured.

[0082] In this embodiment, the sample to be tested is serum, specifically a human serum sample.

[0083] In this embodiment, the mixed volume ratio of the R1 reagent, the R2 reagent and the sample to be tested is 240:60:3.

[0084] The present application utilizes the reaction of glucose kinase with the ATP-Mg complex, the activity of which depends on the amount of the ATP-Mg complex, has a higher specificity for Mg ions, has better stability, and is more accurate in determination; at the same time, the determination method is simple, convenient, and rapid.

[0085] The following is a more specific description of the content of the present application in conjunction with specific embodiments, and further elaboration of the present application, but these embodiments are by no means limiting the present application.

[0086] Example 1

[0087] In this embodiment, the R1 reagent includes phosphate buffer (pH 6.2, 100 mmol / L), glucose (10 mmol / L), ATP (10 mmol / L) and NADP +(0.7mmol / L), Tween80 (surfactant, 5g / L), NaCl (stabilizer, 1g / L) and ProClin300 (preservative, 0.8g / L); R2 reagent includes phosphate buffer (pH 8.5, 100mmol / L), glucose kinase (4KU / L), 6-phosphate glucose dehydrogenase (8KU / L), Tween80 (surfactant, 5g / L), NaCl (stabilizer, 1g / L) and ProClin300 (preservative, 0.8g / L).

[0088] When the reagent of this embodiment is used to measure the sample, the measurement method adopted is the two-point endpoint method, the temperature is 37°C, the usage ratio of R1 reagent: R2 reagent: sample to be measured is 240:60:3, the main / secondary wavelength of measurement is 340 / 700nm, and the R1 reagent is added to the sample to be measured or the standard and incubated at the measurement temperature for 3 minutes. After adding the R2 reagent and incubating for 1 minute, the absorbance of the first point is read, and then the absorbance of the second point is read after 300 seconds.

[0089] The above method and XB method (comparative example) were used to simultaneously measure 20 serum Mg samples. Table 1 is a comparison table of the measured values ​​of this embodiment and the measured values ​​of the conventional XB method. Figure 2 It is made according to the data in Table 1. 2 The correlation coefficient r is 0.9714, and the correlation coefficient r is 0.9856. The two show an excellent correlation, which shows that the present application detects magnesium ions on the basis of more stable reagents and improves the detection accuracy.

[0090] Table 1

[0091]

[0092]

[0093] Example 2

[0094] In this embodiment, the R1 reagent includes triethanolamine-HCl buffer (pH 6.2, 100mmol / L), glucose (10mmol / L), ATP (10mmol / L) and NADP +(0.7mmol / L), Tritonx-100 (surfactant, 5g / L), BSA (stabilizer, 1g / L) and ProClin950 (preservative, 0.8g / L); R2 reagents include triethanolamine-HCl buffer (pH 8.5, 100mmol / L), glucose kinase (4KU / L), 6-phosphate glucose dehydrogenase (8KU / L), Tritonx-100 (surfactant, 5g / L), BSA (stabilizer, 1g / L) and ProClin950 (preservative, 0.8g / L).

[0095] The determination method of this example is the same as that of Example 1. 20 serum Mg samples were simultaneously determined using this method and the XB method. Table 2 is a comparison table of the determination values ​​of Example 2 and the conventional XB method (comparative example). Figure 3 It is made according to the data in Table 2. 2 The correlation coefficient r is 0.9860, which shows an excellent correlation.

[0096] Table 2

[0097]

[0098]

[0099] Example 3

[0100] In this embodiment, the R1 reagent includes Tris-HCl buffer (pH 6.2, 100 mmol / L), glucose (10 mmol / L), ATP (10 mmol / L) and NADP + (0.7mmol / L), Tritonx-405 (surfactant, 5g / L), BSA (stabilizer, 1g / L) and ProClin300 (preservative, 0.8g / L); R2 reagent includes Tris-HCl buffer (pH 8.5, 100mmol / L), glucose kinase (4KU / L), 6-phosphate glucose dehydrogenase (8KU / L), Tritonx-405 (surfactant, 5g / L), BSA (stabilizer, 1g / L) and ProClin300 (preservative, 0.8g / L).

[0101] The determination method of this embodiment is the same as that of Example 1. 20 serum Mg samples were simultaneously determined using this method and the XB method (comparative example). Table 3 is a comparison table of the determination values ​​of this embodiment 3 and the conventional XB method. Figure 4 According to the data in Table 3, the R 2The correlation coefficient r is 0.9792, and the correlation coefficient r is 0.9896, which shows an excellent correlation.

[0102] Table 3

[0103] Comparative ratio determination value (mmol / L) Example Measured value (mmol / L) 0.82 0.86 1.12 1.16 1.13 1.10 0.95 0.97 0.90 0.94 1.02 0.98 1.06 1.12 1.24 1.22 1.21 1.27 1.07 1.11 1.13 1.16 1.01 1.00 0.96 0.98

[0104] Obviously, the embodiments described above are only some embodiments of the present application, rather than all embodiments. The preferred embodiments of the present application are given in the accompanying drawings, but they do not limit the patent scope of the present application. The present application can be implemented in many different forms. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present application more thorough and comprehensive. Although the present application is described in detail with reference to the aforementioned embodiments, for those skilled in the art, it is still possible to modify the technical solutions recorded in the aforementioned specific implementation methods, or to perform equivalent replacement of some of the technical features therein. Any equivalent structure made using the contents of the specification and drawings of this application, directly or indirectly used in other related technical fields, is similarly within the scope of patent protection of this application.

Claims

1. A magnesium ion detection reagent, characterized in that include: R1 reagent and R2 reagent; The components of the R1 reagent include a first buffer, glucose, ATP and NADP + ; The components of the R2 reagent include a second buffer, glucose kinase and 6-phosphate glucose dehydrogenase.

2. The magnesium ion detection reagent according to claim 1, characterized in that The first buffer and the second buffer are independently selected from one of Tris-HCl buffer, HEPES buffer, PIPES buffer, CAPS buffer, glycine buffer, triethanolamine-HCl buffer, citric acid-trisodium citrate buffer, phosphate buffer and potassium hydrogen phthalate-hydrochloric acid buffer.

3. The magnesium ion detection reagent according to claim 1, characterized in that The components of the R1 agent and the R2 agent further independently include a surfactant, a stabilizer and a preservative.

4. The magnesium ion detection reagent according to claim 3, characterized in that The surfactant is selected from at least one of emulsifiers OP, SDS, Tween20, Tween80, Brij35, Brij98, Tritonx-100, Tritonx-405, B66, A90 and 15-S9; The stabilizer is selected from at least one of NaCl, sucrose, polyols, bovine serum albumin and crown ethers; The preservative is selected from one of sodium azide, ProClin series, thimerosal, gentamicin and sodium benzoate.

5. The magnesium ion detection reagent according to claim 4, characterized in that The concentrations of the first buffer and the second buffer are independently 90 to 110 mmol / L; and / or The concentration of glucose is 9-10 mmol / L; and / or The concentration of ATP is 9-10 mmol / L; and / or The NADP + The concentration is 0.5-0.8 mmol / L; and / or The concentration of glucokinase is 3-5 KU / L; and / or The concentration of the 6-phosphate glucose dehydrogenase is 6-10 KU / L; and / or The concentration of the surfactant is 4 to 6 g / L; and / or The concentration of the stabilizer is 0.5 to 1.5 g / L; and / or The concentration of the preservative is 0.5-1.0 g / L.

6. The magnesium ion detection reagent according to any one of claims 1 to 5, characterized in that The pH value of the first buffer solution is 6.0-7.0, and the pH value of the second buffer solution is 8.0-9.

0.

7. A magnesium ion detection kit, characterized in that: The method comprises the magnesium ion detection reagent as described in any one of claims 1 to 6.

8. A method for detecting magnesium ions, characterized in that: The following steps are involved: Step S10, mixing the sample to be tested with the R1 reagent and the R2 reagent, wherein the components of the R1 reagent include a first buffer, glucose, ATP and NADP + ; The components of the R2 reagent include a second buffer, glucokinase and 6-phosphoglucokinase; Step S20, the magnesium ions in the sample to be tested form an ATP-Mg complex with ATP; Step S30, under the catalytic action of glucokinase, the ATP-Mg complex reacts with glucose to obtain glucose-6-phosphate and ADP-Mg, and the ADP-Mg immediately dissociates into magnesium ions and ADP, and the dissociated magnesium ions are circulated for step S20; Step S40, glucose-6-phosphate and NADP + 6-phosphogluconate and NADPH are generated under the action of 6-phosphogluconate dehydrogenase; Step S50, measuring NADPH, and determining the magnesium ion content in the sample to be tested according to the increase rate of NADPH.

9. The method for detecting magnesium ions according to claim 8, characterized in that: The sample to be tested is serum.

10. The method for detecting magnesium ions according to claim 8, characterized in that: The mixed volume ratio of R1 reagent, R2 reagent and the sample to be tested is 240:60:3.

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