5'-nucleotidase diagnosing reagent kit and 5'-nucleotidase activity concentration measurement method

A diagnostic kit and nucleotidase technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, color/spectral characteristic measurement, etc., and can solve problems such as difficulty in popularization and application, strong acid pollution of the environment, and complicated methods

Inactive Publication Date: 2008-01-02
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The isotope substrate method is complicated and has isotope pollution, and requires an isotope analyzer, so it is difficult to practically popularize and apply
Inorganic phosphorus determination method is a method invented in 1925, the accuracy is not good, strong acid pollutes the environment, etc., it is not suitable for popularization and application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] The 5'-nucleotidase diagnostic reagent of this embodiment is a single reagent, comprising:

[0104] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0105] Stabilizer 50mmol / L

[0106] Reduced coenzyme 0.25mmol / L

[0107] Adenosine deaminase 8000U / L

[0108] Glutamate dehydrogenase 150000U / L

[0109] Glutamate oxidase 8000U / L

[0110] α-ketoglutarate 16mmol / L

[0111] Peroxidase 30000U / L

[0112] 4-amino antiarsenic 2mmol / L

[0113] Carbolic acid 10mmol / L

[0114] After the reagents are all dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents; before use, add purified water and reconstitute for use.

[0115] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 505nm, test sub-wavelength 600nm, the volume ratio of the sample to be tested is 1 / 25, and the reaction direction is positive reaction (rising reaction) ), the detection method is the r...

Embodiment 2

[0118] The 5'-nucleotidase diagnostic reagent of this embodiment is a double reagent, comprising:

[0119] Reagent 1

[0120] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0121] Stabilizer 50mmol / L

[0122] Reduced coenzyme 0.25mmol / L

[0123] Glutamate dehydrogenase 120000U / L

[0124] α-ketoglutarate 16mmol / L

[0125] Peroxidase 30000U / L

[0126] 2,4,6-tribromo-3-hydroxy-benzenesulfonic acid 0.2mmol / L

[0127] Reagent 2

[0128] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0129] Buffer 100mmol / L

[0130] Stabilizer 50mmol / L

[0131] Adenosine deaminase 4000U / L

[0132] Glutamate oxidase 8000U / L

[0133] 4-amino antiarsenic 0.6mmol / L

[0134] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0135] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 546nm, test sub-wave...

Embodiment 3

[0137] Embodiment three 5'-nucleotidase diagnostic reagents are three reagents, comprising:

[0138] Reagent 1

[0139] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0140] Stabilizer 50mmol / L

[0141] Reduced coenzyme 0.25mmol / L

[0142] Glutamate dehydrogenase 100000U / L

[0143] α-ketoglutarate 16mmol / L

[0144] 3-methyl-2-benzothiazolone hydrazone 0.6mmol / L

[0145] Reagent 2

[0146] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0147] Stabilizer 50mmol / L

[0148] Peroxidase 40000U / L

[0149] Dimethylaniline 2mmol / L

[0150] Reagent 3

[0151] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0152] Stabilizer 50mmol / L

[0153] Adenosine deaminase 5000U / L

[0154]Glutamate oxidase 8000U / L

[0155] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0156] Set on the automatic biochemical analyzer: temperat...

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PUM

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Abstract

The invention relates to a 5'-nucleotidase diagnosis reagent box of Enzymatic Recycling Method, and the invention also relates to method principle for detecting the solution of 5'-nucleotidase, constitute and component of reagent, which belongs to the technical field of medical checking measurements. The reagent box in the invention can be dry powder state, and used after dissolution; it also can be formulated to be liquid agent for direct usage. The component of reagent box mainly contains: buffering liquid, alpha- ketoglutaric acid, reduction type coenzyme, Adenosine Deiminase EC 3.5.4.4, glutamate dehydrogenase EC 1.4.1.2, EC 1.4.1.3, EC 1.4.1.4, Glutamate oxidase EC 1.4.3.7, EC 1.4.3.11, peroxidase EC 1.11.1.7, reduction type chromogen assembly, stabilizer and anti-interference agent; and the component can be mixed to form single-reagent reagent box, two-reagent reagent box, and three-reagent reagent box; by series of enzymatic reaction, the colorless reduction type chromogen assembly is oxygenated to form color dye, and the content of dye can be measured by visible light analyzer at wavelength of 400-700nm to reflect the concentration of 5'-nucleotidase directly. Comparing with present technique, the invention can be spread easily, and because the produced dye has higher molar extinction coefficient, the sensitivity is high, and the precision is well.

Description

technical field [0001] The invention relates to a 5'-nucleotidase diagnostic kit, and at the same time, the invention also relates to a method for measuring the activity concentration of 5'-nucleotidase, which belongs to the technical field of medical testing. Background technique [0002] Medical research shows that the increase of 5'-nucleotidase (5NT) is mainly seen in obstructive jaundice, and can also be seen in liver cancer and hepatitis. In cholestasis complicated by cholangitis, primary and secondary biliary cirrhosis, and chronic hepatitis, the rate of 5NT elevation is higher than that of alkaline phosphatase; in liver tumors and hepatic granulomas, the sensitivity of 5NT elevation is higher than that of alkali sexual phosphatase. Because there is no physiological increase in 5'-nucleotidase activity, it is more sensitive and specific than alkaline phosphatase for the diagnosis of infantile liver disease and hepatic cholestasis of pregnancy. Therefore, the determi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/78G01N33/50G01N33/52C12Q1/00
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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