A kit and method for detecting 1,5-ag
A 5-AG, kit technology, applied in the field of medical detection, can solve the problems of inability to eliminate endogenous glucose interference, inability to eliminate high-concentration glucose interference, inability to effectively eliminate endogenous glucose interference, etc. The effect of good precision and wide linear range
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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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