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5'-nucleotidase diagnosis reagent kit and 5'-nucleotidase active concentration determination method

A nucleotidase and nucleoside monophosphate technology, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, and color/spectral property measurement, etc.

Inactive Publication Date: 2009-06-24
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

[0042] The 5'-nucleotidase diagnostic reagent of this embodiment is a single reagent, comprising:

[0043] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L

[0044] Stabilizer 500mmol / L

[0045] Reduced coenzyme 0.25mmol / L

[0046] Phosphoenolpyruvate Carboxylase 6000U / L

[0047] Pyruvate dehydrogenase 10000U / L

[0048] Nucleoside monophosphate 8mmol / L

[0049] Oxaloacetate 12mmol / L

[0050] Acetyl-CoA 3mmol / L

[0051] 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.

[0052] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, the volume of the tested 5'-nucleotidase sample and reagent The ratio is 1 / 25, the reaction direction is negative reaction (decreasing reaction), the delay time is about 1 mi...

Embodiment 2

[0055] The 5'-nucleotidase diagnostic reagent of this embodiment is a double reagent, comprising:

[0056] Reagent 1

[0057]Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L

[0058] Stabilizer 50mmol / L

[0059] Reduced coenzyme 0.25mmol / L

[0060] Nucleoside monophosphate 8mmol / L

[0061] Oxaloacetate 12mmol / L

[0062] Acetyl-CoA 3mmol / L

[0063] Reagent 2

[0064] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L

[0065] Stabilizer 500mmol / L

[0066] Phosphoenolpyruvate Carboxylase 6000U / L

[0067] Pyruvate dehydrogenase 10000U / L

[0068] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0069] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, tested 5'-nucleotidase sample and reagent 1, The volume ratio of reagen...

Embodiment 3

[0072] The 5'-nucleotidase diagnostic reagent of the present embodiment is three reagents, comprising:

[0073] Reagent 1

[0074] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L

[0075] Stabilizer 50mmol / L

[0076] Reduced coenzyme 0.25mmol / L

[0077] Nucleoside monophosphate 8mmol / L

[0078] Oxaloacetate 12mmol / L

[0079] Acetyl-CoA 3mmol / L

[0080] Reagent 2

[0081] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L

[0082] Stabilizer 500mmol / L

[0083] Pyruvate dehydrogenase 10000U / L

[0084] Reagent 3

[0085] Tris(carboxymethyl)aminomethane—hydrochloric acid buffer 100mmol / L

[0086] Stabilizer 500mmol / L

[0087] Phosphoenolpyruvate Carboxylase 6000U / L

[0088] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0089] When measuring the concentration of 5'-nucleotidase activity, set it on the automatic biochemical analyzer: tempera...

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PUM

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Abstract

The invention relates to a kit for diagnosing 5'-nucleotidase by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the active concentration of 5'-nucleotidase, and belongs to the technical field of medical inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, nucleoside monophosphate, an oxaloacetic acid, acetylcoenzyme A, phosphoenolpyruvate carboxykinase, pyruvate dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of 5'-nucleotidase.

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): G01N21/31C12Q1/68C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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