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Method for testing oxidative stress biomarkers in blood plasma with low coefficient of variation

A technology of biomarkers and coefficient of variation, applied in the field of oxidative stress biomarkers in testing plasma with low coefficient of variation, can solve the problem of high coefficient of variation, and achieve the effect of improving reliability, reliable measurement method, and increasing possibility

Inactive Publication Date: 2020-04-10
JILIN UNIV
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Problems solved by technology

Clearly this coefficient of variation is too high to be the best way to detect biomarkers of oxidative stress in plasma

Method used

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  • Method for testing oxidative stress biomarkers in blood plasma with low coefficient of variation
  • Method for testing oxidative stress biomarkers in blood plasma with low coefficient of variation
  • Method for testing oxidative stress biomarkers in blood plasma with low coefficient of variation

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Embodiment 1

[0015] The divided plasma was extracted with ethanol / ether (v / v 3:1), and the extraction ratio was plasma:extraction solution=1:20. Example 1 uses 62 μL of plasma and 1238 μL of extract for each sample for extraction. Add the extract to the plasma and shake it well to make the plasma fully react with the extract. Immediately thereafter, centrifuge at 3000 rpm for 10 min at 4°C to precipitate the flocs formed by the reaction. After aspirating the supernatant, centrifuge the supernatant again under the same conditions for 5 min, in order to centrifuge the precipitate sufficiently so as not to affect the measured fluorescence intensity value. The supernatant was loaded into a CORNING 96-well black fluorescent plate (FLATBOTTOM, REF3925), 300 μL was added to each well, and the reading height was 7.00 mm. Fluorescence intensity of the liquid at excitation wavelength / emission wavelength: 320 / 420nm, 360 / 420nm, 400 / 475nm (as shown in Table 1 below).

[0016] In order to facilitate ...

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Abstract

The invention belongs to the technical field of oxidative stress biomarker detection, in particular to a method for testing oxidative stress biomarkers in blood plasma with a low coefficient of variation. The method comprises the following steps: taking a mixture of blood plasma and an extractant with the mixed volume ratio of 1: 20 as a to-be-detected substance solution, taking a supernatant of the to-be-detected substance solution and carrying out fluorescence determination by adopting a fluorescence microplate reader under the conditions of three excitation or emission wavelengths. According to the method, the coefficient of variation of oxidative stress biomarkers in blood plasma is remarkably reduced, the inter-batch coefficient of variation of FlOPs is reduced to be less than 3.6%, and the intra-batch coefficient of variation is reduced to be less than 2.7%, so that a more reliable FlOPs measurement method is provided for future epidemiological research and potential clinical practice.

Description

technical field [0001] The invention belongs to the technical field of detection of oxidative stress biomarkers, and in particular relates to a method for testing oxidative stress biomarkers in blood plasma with a low coefficient of variation. Background technique [0002] Oxidative stress is caused by an imbalance between excess reactive oxygen species and antioxidant capacity in the body. Fluorescent oxidation products (FlOPs) in plasma are a new type of biomarker that can fully reflect the level of oxidative stress in the body. FlOPs can remain stable for up to 48 hours in unpackaged blood samples stored at 4°C. Stored aliquoted blood samples are stable for more than 10 years. Studies have shown that FlOPs are associated with many oxidative stress-related diseases such as cardiovascular disease and chronic kidney disease. [0003] Existing methods for detecting oxidative stress biomarkers in plasma are all based on the extension of Shimasaki's method. That is, the extr...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6419G01N2021/6421G01N2021/6439
Inventor 杨书满张宇铮赵倩倩薛珊珊乔文婧沈雪郭丁杰李彬彬
Owner JILIN UNIV
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