Method for quantitatively detecting uric acid in serum
A quantitative detection and serum technology, which is applied in the field of quantitative detection of uric acid in serum, can solve the problems of unstable protease properties, limited practical application, and harsh conditions for the use of natural enzymes.
Pending Publication Date: 2020-08-14
GUANGDONG NO 2 PROVINCIAL PEOPLES HOSPITAL
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[0002] At present, the method commonly used in clinical serum uric acid detection is mainly the uricase-peroxidase coupling method. The experimental principle is: uric acid can be oxidized by uricase to generate allantoin and hydrogen peroxide, which are Under hydrogen peroxide, 3,5-dichloro-2-hydroxybenzenesulfonic acid (DHBS) and 4-aminoantipyrine (4-AAP) are condensed into red quinone compounds, which have a maximum absorbance at 520nm wavelength, Its absorbance value is directly proportional to the uric acid content; the peroxidase used in this method is a natural protease, such as horseradish peroxidase, which is widely used in various clinical detection methods, but due to the nature of natural protease Instability, low content in organisms, difficult purification and storage, etc., such as easy inactivation under overacid, overalkaline and high temperature conditions, which also make the use conditions of natural enzymes harsh, narrow range Conditions of use limit its practical application
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Embodiment 1
[0042] 1. Experimental equipment preparation: EP tube, sampling gun, sampling gun tip, marker pen, incubation specimen rack, water bath, cuvette, full-wavelength scanner, lotion bottle filled with distilled water;
[0043] Second, the establishment of the standard curve
[0044] Take 6 working standard solutions with a volume of 10 μL, the molar concentrations of uric acid in the 6 working standard solutions are 100 μmol / L, 200 μmol / L, 400 μmol / L, 800 μmol / L, 1200 μmol / L, 1600 μmol / L and respectively Named Standard 1, Standard 2, Standard 3, Standard 4, Standard 5, and Standard 6, the six working standard solutions were respectively operated as follows:
[0045] EP tube 2: 10 μL standard 1 (100 μmol / L uric acid) + 50 μL 1.6 mg / mL uricase + 100 μL: 0.105 mg / MlMCA-Pd NPs + 50 μL 10 mmol / L ABTS 2- ;
[0046] EP tube 3: 10 μL standard 2 (200 μmol / L uric acid) + 50 μL 1.6 mg / mL uricase + 100 μL 0.105 mg / mL MCA-Pd NPs + 50 μL 10 mmol / L ABTS 2- ;
[0047] EP tube 4: 10 μL standard ...
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The invention discloses a method for quantitatively detecting uric acid in serum, which belongs to the technical field of detection of uric acid in serum. The method comprises the following steps: S1,adding uricase into serum to be detected, uniformly performing mixing to obtain a mixed solution A; S2, adding a compound formed by melamine cyanurate and palladium nanoparticles into the mixed solution A to obtain a mixed solution B; S3, adding diammonium 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) into the mixed solution B to obtain a mixed solution C; S4, putting the mixed solution C ina 37 DEG C constant-temperature environment for 15-25 min for incubation, then performing standing for 8-12 min in the room-temperature environment to obtain a mixed solution D; and S5, measuring theabsorbance value of the mixed solution D at 420nm, substituting the measured absorbance value into a calculation formula obtained by the standard curve, and calculating the concentration of uric acid. The concentration of uric acid in serum is quantitatively detected, so that a detection system is simplified, influence factors are reduced, and the reagent cost is lowered.
Description
technical field [0001] The invention belongs to the technical field of uric acid detection in serum, and in particular relates to a method for quantitatively detecting uric acid in serum. Background technique [0002] At present, the method commonly used in clinical serum uric acid detection is mainly the uricase-peroxidase coupling method. The experimental principle is: uric acid can be oxidized by uricase to generate allantoin and hydrogen peroxide, which are Under hydrogen peroxide, 3,5-dichloro-2-hydroxybenzenesulfonic acid (DHBS) and 4-aminoantipyrine (4-AAP) are condensed into red quinone compounds, which have a maximum absorbance at 520nm wavelength, Its absorbance value is directly proportional to the uric acid content; the peroxidase used in this method is a natural protease, such as horseradish peroxidase, which is widely used in various clinical detection methods, but due to the nature of natural protease Instability, low content in organisms, difficult purificat...
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Login to View More IPC IPC(8): G01N21/33G01N1/28
CPCG01N21/33G01N1/28
Inventor 邹瑟音
Owner GUANGDONG NO 2 PROVINCIAL PEOPLES HOSPITAL