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Kit for detection of 1,5-AG and method thereof

A kit and 5-AG technology, applied in the field of medical testing, can solve problems such as the inability to eliminate high-concentration glucose interference, the inability to effectively eliminate endogenous glucose interference, and the inability to eliminate endogenous glucose interference, achieving high accuracy, The effect of good precision and wide linear range

Active Publication Date: 2018-02-16
SINOCARE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, this method cannot effectively eliminate the interference of endogenous glucose
[0026] The HK combined with PK method can eliminate the interference of low concentration of endogenous glucose (1~15mM) in the 1,5-AG test sample, but it cannot eliminate the interference of high concentration of glucose, and there are other interfering substances in the actual sample at this wavelength Under UV absorption
[0027] The above three methods all have limitations. The GOD method and the HK combined with G6PD method cannot eliminate the interference of endogenous glucose. Derived glucose interference

Method used

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  • Kit for detection of 1,5-AG and method thereof
  • Kit for detection of 1,5-AG and method thereof
  • Kit for detection of 1,5-AG and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1～3

[0057] Embodiment 1~3 kit of the present invention

[0058] Prepare the detection kit according to Table 1:

[0059] Table 1

[0060]

[0061] Wherein, the pH value of the MES buffer solution is 5.9, and the pH value of the HEPES buffer solution is 7.2.

Embodiment 4

[0065] Embodiment 4 The anti-interference test of the kit of the present invention to endogenous glucose

[0066] Use a solution similar to human serum matrix: NaCl 0.8%, BSA 0.2%, NaN 3 0.4%, Tris-HCl pH8.0, prepare 1,5-AG samples containing 5, 10, 15, 20, 25, 30mM GLU, and use the sample without glucose as a control, in which the concentration of 1,5-AG is 200 μM. Add the reagent R1 in the kits of Examples 1-3 and Comparative Example 1 of the above scheme to the centrifuge tube, then add 10 μL of the sample to be tested, and react in a 37°C water bath for 5 minutes; then add 75 μL of the corresponding reagent R2, mix well, React in a water bath at 37°C for 5 min, and detect the absorbance at a wavelength of 555 nm. The results are shown in Table 2

[0067] Table 2

[0068]

[0069]

[0070] It can be seen from Table 2 that compared with the HK+PK method disclosed in the prior art, the kits provided in Examples 1 to 3 of the present invention can effectively elimin...

Embodiment 5

[0071] The performance evaluation of embodiment 5 kit of the present invention:

[0072] Dissolve 1,5-AG in a solution similar to human serum matrix (NaCl 0.8%, BSA 0.2%, NaN 3 0.4%, Tris-HCl pH8.0) to make different concentrations of calibrators, using Jiuqiang 1,5-AG detection kit (batch number: 15-1218) and supporting standard concentration to determine the different concentrations of calibrators Values, the concentration of the original calibration standard of 1,5-AG was 25.4μM, 40.4μM, 152.3μM, 247.9μM, 500.6μM, and a 0μM solution was prepared, and a total of 6 concentrations were prepared. Wherein, the glucose concentration is 10 mM.

[0073] Add the reagent R1 in the kit of Example 1 to the centrifuge tube, add 10 μL of calibrator solutions of different concentrations, mix well, incubate at 37°C for 5 minutes, add 75 μL of reagent R2, mix well, react at 37°C for 5 minutes, at 555nm wavelength The absorbance detection was repeated 3 times for each tube, and the standa...

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Abstract

The invention relates to the field of medical testing and discloses a kit for detection of 1,5-AG and a method thereof. According to the kit, by selection of appropriate reagent R1 and reagent R2 andby adoption of HK+G6PD method+ lactate dehydrogenase method, endogenous glucose in a 1,5-AG sample is effectively eliminated, and then the content of 1,5-AG in human serum is detected by the utilization of an oxidase-ultraviolet spectrophotometry method. The test results show that the kit provided by the invention can eliminate the interference of 1-20nM endogenous glucose. In comparison with a detection kit in the prior art, the kit of the invention has advantages of wider application range, high test result accuracy, good precision and wide linear range, and can be widely used for measuringthe content of 1,5-AG in serum in hospitals of different levels, health and prevention departments and research and development institutions of medical biology.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a kit and method for detecting 1,5-AG. Background technique [0002] Interfering substances are the main source of errors in clinical test results. This kind of error is very detrimental to the patient's diagnosis, sometimes leading to very serious consequences. [0003] Diabetes is an important metabolic disease characterized by high blood sugar that endangers human health. Timely and accurate measurement and monitoring of blood sugar can reflect the metabolic status of patients and provide a basis for treatment. Currently, the diagnosis of diabetes includes determination of fasting blood glucose (FBG), oral glucose tolerance test (OGTT), glycosylated hemoglobin (HbA1C) and fructosamine (FA). Both FBG and OGTT tests are affected by changes in diet, drugs, activities, and metabolism in the body; HbA1C and FA tests are not sensitive enough for diabetes screening, and are mainly ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/3103
Inventor 曾丽刘久逢
Owner SINOCARE
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