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5 '-nucleotidase detection kit and preparation method thereof

A technology for detecting kits and nucleotidases, which is applied in biochemical equipment and methods, and the determination/inspection of microorganisms, and can solve the problems of low precision and stability of 5'-nucleotidase detection kits

Active Publication Date: 2016-05-04
JILIN GETEIN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a 5'-nucleotidase detection kit and a preparation method thereof in order to solve the problems of low precision and stability of the existing 5'-nucleotidase detection kits

Method used

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  • 5 '-nucleotidase detection kit and preparation method thereof
  • 5 '-nucleotidase detection kit and preparation method thereof
  • 5 '-nucleotidase detection kit and preparation method thereof

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preparation example Construction

[0034] The present invention also provides a preparation method of a 5'-nucleotidase detection kit, comprising:

[0035] Step 1: Preparation of reagent R1:

[0036] 1. First add deionized water into the reaction vessel, then add buffer I, stir to dissolve, then add potassium dihydrogen phosphate, EDTA 2K, 4-aminoantipyrine, Proclin300 and AES in sequence, stir to dissolve completely, preferably Adjust the pH value of the solution to 7.0-7.7 with an alkaline solution at 20°C. The alkaline solution is preferably KOH, and the concentration of KOH is preferably 4M to obtain a mixed solution;

[0037] 2. Add A90 to the mixed solution obtained in step 1. After there are no insoluble particles in the solution, add MgCl2·6H2O and stir to mix evenly, then use a pipette to absorb TritonX-305 and add it to the above reaction container, stir and mix evenly, and finally Add purine nucleoside phosphorylase, xanthine oxidase and peroxidase, dilute to volume with deionized water, stir evenly...

Embodiment 1

[0043] Preparation of reagent R1: (1L)

[0044] 1) Measure 1L of deionized water with a graduated cylinder, take 0.9L and add it to the beaker, and keep the remaining water for use;

[0045] 2) Weigh 40mmol of PIPES and 80mmol of KOH into the above beaker, stir with a glass rod until completely dissolved;

[0046] 3) Weigh 15mmol of potassium dihydrogen phosphate, 1mmol of EDTA·2K, 1mmol of 4-aminoantipyrine, 0.02ml of Proclin300 and 0.5g of AES into the above-mentioned large beaker, and stir until completely dissolved;

[0047] 4) Adjust the pH value of the above solution to 7.6 with 4M KOH at 20°C;

[0048] 5) Measure 0.1ml of A90 into the above beaker and mix well;

[0049] 6) After the solution in the above beaker has no insoluble particles, weigh 3 mmol of MgCl 2 ·6H 2 O was added to the above-mentioned beaker, and stirred gently;

[0050] 7) Use a pipette to draw 0.1ml of TritonX-305 into the above beaker, stir gently;

[0051] 8) Finally, add 0.2KU of purine nucleos...

Embodiment 2

[0060] preparation:

[0061] Preparation of reagent R1: (1L)

[0062] 1) Measure 1L of deionized water with a graduated cylinder, take 0.9L and add it to the beaker, and keep the remaining water for use;

[0063] 2) Weigh 40mmol of KH 2 PO 4 1. Add 70mmol of KOH into the above beaker, stir with a glass rod until completely dissolved;

[0064] 3) Weigh 2mmol of EDTA·2K, 2mmol of 4-aminoantipyrine, 0.02ml of Proclin300 and 1.0g of AES into the above-mentioned large beaker, and stir until completely dissolved;

[0065] 4) Adjust the pH value of the above solution to 7.6 with 4M KOH at 20°C;

[0066] 5) Measure 0.2ml of A90 into the above beaker and stir well;

[0067] 6) After the solution in the above beaker has no insoluble particles, weigh 6 mmol of MgCl 2 ·6H 2 O was added to the above-mentioned beaker, and stirred gently;

[0068] 7) Use a pipette to draw 0.2ml of TritonX-305 into the above beaker, stir gently;

[0069] 8) Finally, add 0.2KU of purine nucleoside pho...

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Abstract

The present invention provides a 5'-nucleotidase detection kit and a preparation method thereof, belongs to the field of in-vitro diagnostic reagents, and solves the low precision and stability problems of a 5'-nucleotidase detection kit in the prior art. The kit comprises a reagent R1 and a reagent R2. The present invention also provides the preparation method of the 5'-nucleotidase detection kit. The R1 reagent contains a compound stabilizer comprising TritonX-305 and AES, the compound stabilizer can improve the stability of the reagent, and also can improve the dispersion of the reagent, reduce the absorbance difference between two times of blank calibration, and improve the kit precision; A90 contained in the R1 can effectively prevent precipitation of magnesium phosphate, expensive 18-crown ether-6 is replaced, the stability of the reagent is improved, reagent blank absorbance change rate is reduced, and the cost is reduced greatly; by addition of glycine and EDTA. 2K into the reagent R2, detection sensitivity and precision of the kit can be improved, and by addition of the glycine, reagent blank absorbance is reduced.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to a 5'-nucleotidase detection kit and a preparation method thereof. Background technique [0002] 5'-Nucleotidase (5'-Nucleotidase, 5'-NT) determination is mainly used for the diagnosis of hepatobiliary system diseases and the differential diagnosis of bone diseases. Increased serum 5'-NT activity is mainly seen in diseases of the hepatobiliary system, such as obstructive jaundice, liver cancer, hepatitis, etc., and its activity can be increased by 2-6 times, and it is positively correlated with the severity of the disease, and its activity change is consistent with ALP. However, in diseases of the skeletal system, such as tumor metastasis, osteitis deformans, rickets, hyperparathyroidism, etc., ALP activity is usually elevated, while 5'-NT is normal. Therefore, the simultaneous determination of ALP and 5'-NT is helpful for the differential diagnosis of hepatobiliary an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48C12Q1/44C12Q1/28C12Q1/26
CPCC12Q1/26C12Q1/28C12Q1/44C12Q1/48C12Q2326/96
Inventor 苏恩本徐李鹏刘玲张晓杨
Owner JILIN GETEIN BIOTECH CO LTD
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