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

A technology of oxalic acid and reagents, which is applied in the field 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 effect of simple steps, low cost, and high sensitivity

Active Publication Date: 2015-03-11
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 content of oxalic acid in urine and blood
  • Reagent, kit and method for detecting content of oxalic acid in urine and blood
  • Reagent, kit and method for detecting content of oxalic acid in urine and blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Step 101: preparing an oxalic acid assay reagent, adding 10umol Amplex red, 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 th...

Embodiment 2

[0068] Determination of urinary oxalate concentration

[0069] Step 201: Prepare oxalic acid determination reagent, add 10umol Amplex red, 1000U HRP and 100U oxalate oxidase to every liter of buffer solution with pH value of 5.4 to obtain oxalic acid determination 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 meth...

Embodiment 3

[0081] Step 301: preparing reagents for oxalic acid determination, adding 1 umol Amplex red, 100 U HRP and 10 U oxalate oxidase to each liter of buffer solution with a pH value of 4.0 to obtain reagents for determining oxalic acid.

[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: Th...

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Abstract

The invention discloses a reagent, a kit and a method for detecting the content of oxalic acid in urine and blood and relates to the field of scientific research and detection for medical oxalic acid. The reagent comprises a buffer solution of which the pH value is 4.0-7.5, Amplex red fluorescent dye of which the concentration is 1-50[mu]mol / L, horse radish peroxidase (HRP) of which the concentration is 0.1-2U / ml and oxalate oxidase of which the concentration is 0.01-1.0U / ML. The method comprises the following steps of collecting and purifying a sample to be detected to obtain a purified liquid to be detected; dividing the liquid to be detected into two parts; preparing the liquid to be detected into a calibrated liquid to be detected; dividing the liquid to be detected of each part into two groups, namely a determining group and a comparing group; adding an oxalic acid determining reagent in the determining group to obtain a fluorescent group to be detected; adding an oxalic acid background reagent in the comparing group to obtain a determined fluorescent value after comparing the groups to be detected, and obtaining the detection limit of the oxalic acid, namely 1.5-3mol / L. The reagent, the kit and the method disclosed by the invention, is used for scientific research and clinical medicine, and high throughout detection can be realized; the method is high in sensitivity and better in 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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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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