Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'nucleotidase
A diagnostic kit and nucleotidase technology are applied in the field of determining 5′-nucleotidase activity and 5′-nucleotidase diagnostic kits, which can solve the problem of being unsuitable for popularization and application, environment polluted by strong acid, and difficult to popularize. Application and other issues, to achieve the effect of low test cost, accurate test results, and easy promotion and application
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Embodiment 1
[0062] Embodiment one (single agent)
[0063] The 5'-nucleotidase diagnostic kit of the present embodiment comprises:
[0064] Buffer 80mmol / l
[0065] Uridine monophosphate 2mmol / l
[0066] Oxidized coenzyme 2mmo / l
[0067] Uridine nuclease 5000U / l
[0068] Ribose 1-dehydrogenase 5000U / l
[0069] Stabilizer 50% (total volume)
[0070] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the tested 5'-nucleotidase sample to the reagent is 1 / 25, The reaction direction is positive reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.
[0071] After the sample and reagents are added and allowed to mix, the following reactions occur:
[0072] Uridine monophosphate + water 5 nucleotidase Uridine + Phosphate
[0073] Uridine + water uridine nuclease ribose + uracil
[0074] Ribose + oxidized coenzyme...
Embodiment 2
[0077] Embodiment two (two doses)
[0078] The 5'-nucleotidase diagnostic reagent of the present embodiment has:
[0079] Reagent I
[0080] Buffer 100mmol / l
[0081] Oxidized coenzyme 4mmo / l
[0082] Uridine nuclease 8000U / l
[0083] Ribose 1-dehydrogenase 8000U / l
[0084] Stabilizer 50% (total volume)
[0085] Reagent II
[0086] Buffer 100mmol / l
[0087] Uridine monophosphate 6mmol / l
[0088] Stabilizer 20mmol / l
[0089] When measuring 5'-nucleotidase activity, the temperature is controlled at 30°C, the reaction time is 15 minutes, the main wavelength of the test is 340nm, the secondary wavelength of the test is above 405nm, and the volume ratio of the tested 5'-nucleotidase sample to the reagent is 1 / 25, the reaction direction is positive reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.
[0090] The specific measurement steps are:
[0091] Uridine monophosphate + water 5 nucleotidase Uridine + Phosp...
Embodiment 3
[0096] Embodiment three (three doses)
[0097] The 5′-nucleotidase diagnostic reagent of the present embodiment is three doses, including:
[0098] Reagent I
[0099] Buffer 120mmol / l
[0100] Oxidized coenzyme 6mmo / l
[0101] Stabilizer 20mmol / l
[0102] Reagent II
[0103] Buffer 120mmol / l
[0104] Uridine nuclease 10000U / l
[0105] Ribose 1-dehydrogenase 10000U / l
[0106] Stabilizer 50% (total volume)
[0107] Reagent III
[0108] Buffer 120mmol / l
[0109] Uridine monophosphate 10mmol / l
[0110] Stabilizer 20mmol / l
[0111] Set on the automatic biochemical analyzer: temperature 25°C, reaction time 20 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the tested 5′-nucleotidase sample to the reagent is 1 / 25, The reaction direction is positive reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.
[0112] The specific measurement steps are:
[0113] Uridine monophosphat...
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