Method for rapidly detecting creatinine

Abstract

The invention relates to the technical field of medical detection, in particular to a method for rapidly detecting creatinine, which comprises the following steps: adding a reagent 1, uniformly mixing with a sample, starting timing waiting for 5 minutes, adding a reagent 2, uniformly mixing, starting creatinine reaction, and at the moment, detecting the chemical reaction change rate of a creatinine mixture irregularly. Timing is started from the completion of addition of the reagent 2, the reaction light absorption intensity DA is recorded after A time, then the reaction rate Dn of fixed adjacent time within the time interval N is detected, the fastest rate Kmax of the marked light absorption rate is searched, the creatinine concentration is converted through Kmax, in the reaction process of the creatinase, under the condition of constant temperature, the rate of the creatinase is in direct proportion to the sample concentration, and the creatinine concentration is calculated through Kmax. Therefore, the reaction rate is converted through signal measurement and calculation without waiting for complete reaction of the creatinine, so that the concentration of the creatinine is measured and calculated, the creatinine is rapidly detected, the reaction time is shortened, the user experience is improved, and the user time is saved.

Description

Technical Field

[0001] The invention relates to the technical field of medical detection, and in particular to a method for rapidly detecting creatinine. Background Art

[0002] In the field of in vitro diagnostic analytical instruments and equipment, the principle of urine creatinine detection is that creatine guanyl hydrolase, sarcosine oxidase, creatinine amidohydrolase and peroxidase react with creatinine in the sample by enzyme linkage reaction, and finally develop color through TRINDER'S reaction. The intensity of color development is proportional to the creatinine content in the sample, and the creatinine content in the sample is calculated by comparison with the standard.

[0003] According to the requirements of relevant reagents, reagent 1 first reacts with the sample for 5 minutes, and then reagent 2 is added to react for 5 minutes. The entire reaction time for a single sample is 10 minutes. Adding the sampling, adding, and cleaning time, the entire test time exceeds 10 minutes, which is relatively slow.

[0004] It can be seen in the instructions of the creatinine test kits currently on the market that they are basically implemented according to the above-mentioned reaction process, and the test time is generally the same, all more than 10 minutes. The detection time is slow and the efficiency is low, resulting in patients having to wait for a long time after completing the sampling before they can get their test reports, which affects the user experience and wastes the user's time. Summary of the invention

[0005] The purpose of the present invention is to provide a method for quickly detecting creatinine, which solves the problem that when the existing creatinine kit is used for creatinine detection, the detection time is slow, the efficiency is low, the user experience is affected, and the user's time is wasted.

[0006] To achieve the above object, the present invention provides a method for rapid detection of creatinine, comprising the following steps:

[0007] Add reagent 1 and the sample to be tested into the reaction cup and mix well to form mixture 1;

[0008] Let it stand for a set time, waiting for the endogenous creatine and creatine reaction to finish;

[0009] Add reagent 2, mix with mixture 1 to form mixture 2, and start the reaction;

[0010] When the urine is left to stand for a while, record the time A and the intensity of the urine creatinine reaction D at point A. A ;

[0011] Continue to monitor the urine creatinine reaction value until time N, and record the urine creatinine reaction intensity D from point A to point N B -D n Multiple point values ​​of

[0012] According to D between points A and NA -D n The value between the two is used to find out the maximum reaction rate Kmax of urine creatinine in this process and convert it into creatinine concentration.

[0013] When the urine is left to stand for a while, record the time A and the intensity of the urine creatinine reaction D at point A. A , the steps further include:

[0014] The time at point A is within 20 seconds.

[0015] Among them, the urine creatinine reaction value is continuously monitored until time N, and the reaction intensity D of urine creatinine from point A to point N is recorded. B -D n The step further comprises:

[0016] The time for point N is within 180 seconds.

[0017] Among them, the step of standing for a set time to wait for the endogenous creatine and creatine reaction to end, further comprises:

[0018] Let sit for 5 minutes.

[0019] Wherein, the reagent 1 and the sample to be tested are added into the reaction cup and mixed to form a mixture 1, and the step further includes:

[0020] The sampling needle samples reagent 1 and adds it to the reaction cup;

[0021] The sampling needle samples the sample to be tested, adds it into the reaction cup, and mixes it to form a mixture 1.

[0022] The method for rapid detection of creatinine of the present invention comprises the following steps: firstly, endogenous creatine is eliminated, and no color reaction is generated in this process. After adding reagent 1 and mixing with the sample, a timing waiting is started, and the timing waiting is 5 minutes. Then, creatinine enzyme is added, and creatinine generates H2O2 under the action of creatinine enzyme, creatinase and sarcosine oxidase. Catalase is inactivated under the action of a catalase inhibitor, so that the generated H2O2 is colored in the Trinder reaction. That is, after adding reagent 2 and mixing, a creatinine reaction is started. At this time, the chemical reaction change rate of the creatinine mixture is detected irregularly. The timing is started from the completion of adding reagent 2. After a time A, the reaction absorbance intensity D is recorded. A, then detect the reaction rate Dn of the fixed adjacent time within the N time interval, and find the fastest rate Kmax of the labeled absorbance rate, where the time at point A is controlled within 20 seconds, and the time at point N is controlled within 180 seconds. At this time, the entire reaction of creatinine is completed, and the cleaning process and test result calculation process are entered. The creatinine concentration is converted through Kmax, and the detection instrument enters the next test preparation state. During the reaction process, under the condition of constant temperature, the rate of creatininase is proportional to the sample concentration, so the reaction rate is converted through signal measurement, and there is no need to wait for the complete reaction of creatinine, so the creatinine concentration can be calculated, creatinine can be detected quickly, the reaction time can be shortened, and the user experience can be improved and the user time can be saved. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art are briefly introduced below.

[0024] Figure 1 4 is a flow chart of a method for rapid detection of creatinine according to a first embodiment of the present invention.

[0025] Figure 2 4 is a signal value curve diagram of the creatinine reaction according to the first embodiment of the present invention.

[0026] Figure 3 4 is a graph showing the creatinine reaction rate according to the first embodiment of the present invention.

[0027] Figure 4 4 is a graph showing the relationship between the maximum creatinine reaction rate and the creatinine concentration according to the first embodiment of the present invention.

[0028] Figure 5 1 is a step diagram of a method for rapid detection of creatinine according to a first embodiment of the present invention. DETAILED DESCRIPTION

[0029] Embodiments of the present invention are described in detail below. Examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, but should not be construed as limiting the present invention.

[0030] The first embodiment of the present application is:

[0031] See also Figures 1 to 5 ,in, Figure 1 4 is a flow chart of a method for rapid detection of creatinine according to a first embodiment of the present invention. Figure 2 4 is a signal value curve diagram of the creatinine reaction according to the first embodiment of the present invention. Figure 3 4 is a graph showing the creatinine reaction rate according to the first embodiment of the present invention. Figure 4 4 is a graph showing the relationship between the maximum creatinine reaction rate and the creatinine concentration according to the first embodiment of the present invention. Figure 51 is a step diagram of a method for rapid detection of creatinine according to a first embodiment of the present invention.

[0032] The present invention provides a method for rapidly detecting creatinine, comprising the following steps:

[0033] S101: adding reagent 1 and the sample to be tested into a reaction cup and mixing them to form a mixture 1;

[0034] S102: Standing for a set time, waiting for the endogenous creatine and creatine reaction to end;

[0035] S103: adding reagent 2, mixing with mixture 1 to form mixture 2, and starting the reaction;

[0036] S104: When the urine starts to rest, record the time A and the reaction intensity D of urine creatinine at point A. A ;

[0037] S105: Continue to monitor the urine creatinine reaction value until time N, and record the urine creatinine reaction intensity D from point A to point N B -D n Multiple point values ​​of

[0038] S106: According to the D between points A and N A -D n The value between the two is used to find out the maximum reaction rate Kmax of urine creatinine in this process and convert it into creatinine concentration.

[0039] Specifically, the first step: the sampling needle samples reagent 1 and adds it to the reaction cup, wherein reagent 1: phosphate buffer 50mmol / L, creatinase 2KU / L, sarcosine oxidase 8KU / L, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine sodium salt 1.2mmol / L; the second step: the sampling needle samples the sample to be tested, adds it to the reaction cup, and mixes it to form a mixture 1; the third step: let it stand for 5 minutes, waiting for the endogenous creatine and sarcosine reaction to end, to avoid affecting the subsequent reaction 2. This step of the reaction No color reaction occurs; Step 4: Add reagent 2, mix with mixture 1 to form mixture 2, and start the reaction. This step is a color reaction, where reagent 2: phosphate buffer 50mmol / L, creatinine 30KU / L, peroxidase 7.5KU / L, 4-aminoantipyrine 1mmol / L, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-toluidine sodium salt 0.3mmol / L; Step 5: When starting to stand, record the time A, and record the reaction intensity D of urine creatinine at point A A ; Step 6: Continue to monitor the urine creatinine reaction value until time N, and record the urine creatinine reaction intensity D from point A to point N B ......D n Equal point values; Step 7: According to D between points A to NA ~D n The value between the two is used to find the maximum reaction rate Kmax of urine creatinine in this process, and convert it to creatinine concentration. The formula is: Creatinine concentration = A*Kmax^3+B*Kmax^2+C*Kmax+D; Step 8: At this point, the creatinine reaction is completed, and the urine creatinine concentration is converted according to the maximum reaction rate Kmax. The overall idea of ​​creatinine detection is that during the reaction process, when the temperature is constant, the rate of creatininase is proportional to the sample concentration. Therefore, the reaction rate is converted through signal measurement, and there is no need to wait for the creatinine to react completely, thereby calculating the creatinine concentration.

[0040] The difference from conventional creatinine tests on the market:

[0041] The time of reaction 2 is determined by the maximum reaction rate according to the creatinine reaction rate. Urine creatinine detection is generally controlled within 3 minutes, while the urine creatinine detection on the market has a fixed reaction time of 5 minutes, and the endpoint value is taken. In practical applications, the present invention can shorten the overall urine creatinine detection time, is faster, improves user experience and saves user time.

[0042] What is disclosed above is only one or more preferred embodiments of the present application, and cannot be used to limit the scope of rights of the present application. Ordinary technicians in this field can understand that all or part of the processes of implementing the above embodiments and equivalent changes made according to the claims of the present application are still within the scope covered by the present application.

Claims

1. A method for rapid detection of creatinine, characterized in that: The following steps are involved: Add reagent 1 and the sample to be tested into the reaction cup and mix well to form mixture 1; Let it stand for a set time, waiting for the endogenous creatine and creatine reaction to finish; Add reagent 2, mix with mixture 1 to form mixture 2, and start the reaction; When the urine is left to stand for a while, record the time A and the intensity of the urine creatinine reaction D at point A. A ; Continue to monitor the urine creatinine reaction value until time N, and record the urine creatinine reaction intensity D from point A to point N B -D n Multiple point values ​​of According to D between points A and N A -D n The value between the two is used to find out the maximum reaction rate Kmax of urine creatinine in this process and convert it into creatinine concentration.

2. The method for rapid detection of creatinine as claimed in claim 1, characterized in that: When the urine is left to stand for a while, record the time A and the intensity of the urine creatinine reaction D at point A. A , the steps further include: The time at point A is within 20 seconds.

3. The method for rapid detection of creatinine as claimed in claim 1, characterized in that: Continue to monitor the urine creatinine reaction value until time N, and record the urine creatinine reaction intensity D from point A to point N B -D n The steps further include: The time for point N is within 180 seconds.

4. The method for rapid detection of creatinine as claimed in claim 1, characterized in that: The step of standing for a set time to wait for the endogenous creatine and creatine to react is also as follows: Let sit for 5 minutes.

5. The method for rapid detection of creatinine as claimed in claim 1, characterized in that: Adding reagent 1 and the sample to be tested into a reaction cup and mixing them to form a mixture 1, the step further includes: The sampling needle samples reagent 1 and adds it to the reaction cup; The sampling needle samples the sample to be tested, adds it into the reaction cup, and mixes it to form a mixture 1.

Images