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Reagent, kit and method for detecting oxalic acid content in urine and blood

A technology for middle oxalic acid and oxalic acid, applied in the fields of scientific research and medical oxalic acid detection, can solve the problems of low content of active ingredients, poor reliability and low sensitivity, and achieve the effects of simple steps, low cost and high sensitivity

Active Publication Date: 2017-08-25
WUHAN KANGFUDE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the small molecular weight of oxalic acid, it is easy to fall off after reacting with the monoclonal antibody. The antibody-antigen-enzyme-labeled antibody complex formed after thorough washing has a low content of active ingredients. The detection sensitivity of this method is low and the reliability is poor.

Method used

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  • Reagent, kit and method for detecting oxalic acid content in urine and blood
  • Reagent, kit and method for detecting oxalic acid content in urine and blood
  • Reagent, kit and method for detecting oxalic acid content in urine and blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Step 101 : prepare an oxalic acid assay reagent, add 10umol Amplexred, 1000U HRP and 100U oxalate oxidase to each liter of buffer solution with a pH value of 5.4 to obtain an oxalate assay reagent.

[0057] To prepare oxalic acid background reagent, add 10umol Amplex red and 1000U HRP to each liter of buffer solution with a pH value of 5.4 to obtain oxalic acid background reagent (except that oxalic acid oxidase is not added to the oxalic acid background reagent, other components and concentrations are determined with oxalic acid same reagent).

[0058] To prepare a standard oxalic acid reagent, add 10 mmol oxalic acid to every liter of buffer solution with a pH value of 5.4 to obtain a standard oxalic acid reagent.

[0059] Step 102: Take more than 20 normal rat sera to be tested and mix them in equal proportions to obtain a standard serum pool. According to the processing method of S201, the purified test solution is obtained.

[0060]Step 103: Take 350ul of the pur...

Embodiment 2

[0068] Determination of urinary oxalate concentration

[0069] Step 201: preparing an oxalic acid assay reagent, adding 10umol Amplexred, 1000U HRP and 100U oxalate oxidase to each liter of buffer solution with a pH value of 5.4 to obtain an oxalate assay reagent.

[0070] To prepare oxalic acid background reagent, add 10umol Amplex red and 1000U HRP to each liter of buffer solution with a pH value of 5.4 to obtain oxalic acid background reagent (except that oxalic acid oxidase is not added to the oxalic acid background reagent, other components and concentrations are determined with oxalic acid same reagent).

[0071] To prepare a standard oxalic acid reagent, add 10 mmol oxalic acid to every liter of buffer solution with a pH value of 5.4 to obtain a standard oxalic acid reagent.

[0072] Step 202: Take the urine of normal test rats whose quantity is greater than 20 and mix them in equal proportions to obtain a standard urine pool. According to the processing method of S20...

Embodiment 3

[0081] Step 301 : prepare an oxalic acid determination reagent, add 1 umol Amplexred, 100 U HRP and 10 U oxalate oxidase to each liter of buffer solution with a pH value of 4.0 to obtain an oxalic acid determination reagent.

[0082] Prepare oxalic acid background reagent, add 1umol Amplex red and 100U HRP to each liter of buffer solution with a pH value of 4.0, and obtain oxalic acid background reagent (except that oxalic acid oxidase is not added to the oxalic acid background reagent, other components and concentrations are determined with oxalic acid same reagent).

[0083] To prepare a standard oxalic acid reagent, add 10 mmol oxalic acid to every liter of buffer solution with a pH value of 4.0 to obtain a standard oxalic acid reagent.

[0084] Step 302: This step is the same as step 102 in Embodiment 1.

[0085] Step 303: This step is the same as step 103 in Embodiment 1.

[0086] Step 304: This step is the same as step 104 in Embodiment 1.

[0087] Step 305: This step...

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Abstract

The invention discloses a reagent, kit and method for detecting oxalic acid content in urine and blood, and relates to the field of scientific research and medical oxalic acid detection. The reagent includes a buffer solution with a pH value of 4.0-7.5 and a concentration of 1-50umol Amplex red fluorescent dye per L, horseradish peroxidase HRP with a concentration of 0.1~2U / ml and oxalate oxidase with a concentration of 0.01~1.0U / ml, the method includes the following steps: collecting and purifying the sample to be tested, After obtaining the purified test solution, divide it into several parts, prepare the standard test solution, and divide each part of the standard test solution into two groups: the measurement group and the comparison group, and add the oxalic acid determination reagent to the test group to obtain the fluorescence test group; Add oxalic acid background reagent to the comparison group to obtain the comparison test group, then measure the fluorescence value, and the detection limit of oxalic acid is 1.5-3nmol / L. The invention can realize high-throughput detection and can be used in scientific research and clinical medical detection. The method has high sensitivity and good reliability.

Description

technical field [0001] The invention relates to the field of scientific research and medical oxalic acid detection, in particular to a reagent, kit and method for detecting oxalic acid content in urine and blood. Background technique [0002] Ox (oxalic acid, oxalic acid) is a molecular formula C 2 o 4 h 2 , A binary organic acid with a molecular weight of 90.04. Oxalic acid in the human body includes exogenous oxalic acid and endogenous oxalic acid. Exogenous oxalic acid refers to oxalic acid ingested through diet. Endogenous oxalic acid refers to oxalic acid produced by the human liver itself. It is basically the same as the content of endogenous oxalic acid. Oxalic acid is the final product in the human body and cannot be further metabolized and degraded. It can only be excreted through the blood circulation to the urinary system. [0003] When the oxalic acid content in the blood and urine in the human body is too high, it is easy to form insoluble calcium oxalate c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 刘海峰李娜姚琦周健文李春艳贺晓霞
Owner WUHAN KANGFUDE BIOTECH CO LTD
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