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Method for determining stability of diagnostic reagent of adenosine deaminase by improving coupling enzymatic reaction

A technology of adenosine deaminase and enzymatic reaction, applied in the field of improving the stability of adenosine deaminase diagnostic reagents by coupled enzymatic reaction, which can solve the waste of reagents, poor stability of PNP and XOD enzymes, and difficult determination Results and other issues to achieve the effect of reducing waste and improving stability

Inactive Publication Date: 2009-08-26
AILEX TECH GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the reagents for the determination of adenosine deaminase by coupling enzymatic reactions are basically designed according to the combined use of inosine-coupled PNP and XOD enzymatic reactions and Trider reactions. Due to the poor stability of PNP and XOD enzymes, As a result, even if the reagents are stored in a refrigerated environment, it is not easy to obtain reliable measurement results, and it is easy to cause waste of reagents

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] According to the following ratio, prepare the basic reagent and divide it into 8 parts:

[0038] Reagent 1 (R1) PH7.0

[0039] Tris-HCl 50mmol / l

[0040] 4AAP 2mmol / l

[0041] PNP 0.1u / ml

[0042] XOD 0.2u / ml

[0043] Peroxidase 0.6u / ml

[0044] Reagent 2 (R2) PH4.0

[0045] Tris-HCl 50mmol / l

[0046] Adenosine 10mmol / l

[0047] EHPST 2mmol / l

[0048] Then in the reagent 1 (R1) of 7 parts of the basic reagent, add the following stabilizers respectively, and make the following content:

[0049] EDTA-Ca 100mmol / L

[0050] EDTA-Fe 100mmol / L

[0051] Sodium molybdate 100mmol / L

[0052] Ammonium molybdate 100mmol / L

[0053] Sodium glutamate 100mmol / L

[0054] Bovine serum albumin 10g / L

[0055] Superoxide dismutase (SOD) 10.0KU / L

[0056] Seal the reagent and store it at 4°C, and test the adenosine deaminase quality control product before putting it in, and after putting it in for 1 month and 3 months. Set the measured value of the quality control before bein...

Embodiment 2

[0078] Test method and basic reagent are identical with embodiment 1,

[0079] The addition amount of stabilizer is as follows:

[0080] EDTA-Ca 10mmol / L

[0081] EDTA-Fe 10mmol / L

[0082] Sodium molybdate 5mmol / L

[0083] Ammonium molybdate 5mmol / L

[0084] Sodium glutamate 5mmol / L

[0085] Bovine serum albumin 8g / L

[0086] Superoxide dismutase (SOD) 0.5KU / L

[0087] The stability test results are shown in Table 2:

[0088] Table 2

[0089] stabilizer

[0090] It can be seen from the results in Table 2 that after adding the stabilizer according to the above ratio, the stability of the reagent can be significantly improved.

Embodiment 3

[0092] Test method and basic reagent are identical with embodiment 1,

[0093] The addition amount of stabilizer is as follows:

[0094] EDTA-Ca 50mmol / L

[0095] EDTA-Fe 60mmol / L

[0096] Sodium molybdate 55mmol / L

[0097] Ammonium molybdate 45mmol / L

[0098] Sodium glutamate 45mmol / L

[0099] Bovine serum albumin 9g / L

[0100] Superoxide dismutase (SOD) 6KU / L

[0101] The stability test results are shown in Table 3:

[0102] table 3

[0103] stabilizer

Determination before putting in (%)

Put in 4 degrees January (%)

Put in 4 degrees March (%)

No added substance

100

52

31

EDTA-Ca(II)

100

99

99

EDTA-Fe(III)

100

100

99

Sodium molybdate

100

99

97

Ammonium molybdate

100

98

97

sodium glutamate

100

98

97

bovine serum albumin

100

99

98

Superoxide dismutase

100

99...

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Abstract

The invention provides a method for determining the stability of a diagnostic reagent of adenosine deaminase by improving a coupling enzymatic reaction, which comprises the steps of adding different stabilizing agents in the diagnostic reagent of the adenosine deaminase, wherein the stabilizing agents comprise 10 to 100 mmol / L of calcium ethylene diamine tetracetate, 10 to 100 mmol / L of ferric ethylene diamine tetracetate, 5 to 100 mmol / L of sodium molybdate, 5 to 100 mmol / L of ammonium molybdate, 5 to 100 mmol / L of sodium glutamate, 8 to 10g / L of bovine serum albumin, and 0.5 to 10.0 KU / L of superoxide dismutase. A coupling enzymatic reaction reagent for determining the adenosine deaminase has higher stability by adding any one of the stabilizing agents into the reagent and maintaining the stabilizing agents within the concentration range; the method, which can be widely applied to clinical in vitro diagnosis in vitro and reduces the waste of the reagents.

Description

technical field [0001] The invention relates to a method for improving the stability of adenosine deaminase diagnostic reagent for coupling enzymatic reaction determination. Background technique [0002] Adenosine deaminase (ADA) is a nucleotide aminohydrolase and an important enzyme in purine nucleoside metabolism. It is widely distributed in various tissues of the human body, and its content is relatively high in thymus, spleen and other lymphoid tissues. It is most abundant in red blood cells and D lymphocytes. The increase of ADA is a response of D lymphocytes to local stimulation of some special lesions, which is closely related to the proliferation, differentiation and number of D lymphocytes. Therefore, the detection of ADA is concerned in the research of the diagnosis, treatment and pathogenesis of certain diseases. Among hepatobiliary diseases, the positive rate (70-88.8%) and the most significant increase of serum ADA activity in patients with liver cirrhosis (2-...

Claims

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Application Information

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IPC IPC(8): C12Q1/527
Inventor 金敏崔鹏飞
Owner AILEX TECH GRP CO LTD
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