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A kind of detection reagent of lipoprotein-related phospholipase a2, preparation method and application thereof

A technology for detecting reagents and phospholipases, which is applied in the field of clinical medical parameter detection, can solve the problems of reducing measurement sensitivity, increasing background measurement values, instability, etc., and achieve components that avoid unstable measurement results and improve detectability, The effect of reducing the background value

Active Publication Date: 2022-07-26
浙江伊利康生物技术有限公司
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Problems solved by technology

However, there are substrates that are difficult to dissolve in water, or are soluble in water under the condition of cosolvent but are naturally hydrolyzed to cause instability, the reagent cannot be stored for a long time, and the measured value of the background is increased to reduce the sensitivity of the determination; Enzymes can also hydrolyze the added substrate analogs with 4-nitrophenol groups, resulting in inaccurate measurement results; the end point of the enzyme-catalyzed reaction cannot be determined, and can only be determined by the time point of the measurement; the reaction product 4-nitrophenol The detectability of phenol in the reaction system is not high, and it is easily interfered by other substances

Method used

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  • A kind of detection reagent of lipoprotein-related phospholipase a2, preparation method and application thereof
  • A kind of detection reagent of lipoprotein-related phospholipase a2, preparation method and application thereof
  • A kind of detection reagent of lipoprotein-related phospholipase a2, preparation method and application thereof

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preparation example Construction

[0085] The present invention also provides a preparation method of a detection reagent for lipoprotein-related phospholipase A2, the preparation method comprising the following steps:

[0086] Prepare the volume of the first reagent as needed, calculate and weigh the ingredients according to the following proportions: demulsifier 100mg / L~220mg / L, cosolvent 10wt%~20wt%, buffer 30mmol / L~100mmol / L, chelating agent 10mmol / L~30mmol / L, organic acid 5mmol / L~60mmol / L, inorganic salt 80mmol / L~120mmol / L, dissolve in corresponding volume of deionized water, and adjust the pH value with 2mol / L~5mol / L hydrochloric acid To 4.0~7.6, get the first reagent;

[0087] Prepare the volume of the second reagent as needed, calculate and weigh the ingredients according to the following ratio: 0.5wt% to 2.7wt% of the substrate, 0.01wt% to 0.5wt% of the emulsifier, and then mix the substrate and the emulsifier at a certain temperature Mixing, adding deionized water under stirring conditions, and homo...

Embodiment 1-6

[0105] The detection reagent for lipoprotein-related phospholipase A2 of the present invention is prepared according to the following steps and conditions.

[0106] Prepare the volume of the first reagent as required, calculate and weigh the ingredients of the following components according to the specific components and proportions shown in Table 1: demulsifier, cosolvent, buffer, chelating agent, organic acid, inorganic salt, Be dissolved in the deionized water of the corresponding volume, embodiment 1 uses the hydrochloric acid that concentration is 2mol / L to adjust pH to 7.6, embodiment 2 uses the hydrochloric acid that concentration is 4.5mol / L to adjust pH to 4.0, embodiment 3 uses concentration to be The hydrochloric acid of 3mol / L is adjusted pH to 5.5, embodiment 4 uses the hydrochloric acid that concentration is 4mol / L to adjust pH to 6.2, embodiment 5 uses the hydrochloric acid that concentration is 3.5mol / L pH is adjusted to 5, embodiment 6 uses The hydrochloric ac...

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Abstract

The invention discloses a detection reagent, preparation method and application of lipoprotein-related phospholipase A2. The detection reagent includes a first reagent, a second reagent and a third reagent, the first detection reagent includes a demulsifier, the second detection reagent includes a substrate and an emulsifier, and the third detection reagent includes an enzyme inactivator and an alkali metal salt. The preparation method of the detection reagent includes an emulsification step. The application method of the detection reagent includes the step of mixing and shaking the first reagent and the second reagent. The present invention avoids the hydrolysis of the substrate during the storage process of the detection reagent by changing the existing state of the substrate in the detection reagent, and can quickly dissolve in the detection system to participate in the enzyme catalysis reaction during detection, thereby reducing the amount of other enzymes in the sample to be tested. It can provide the end point of the enzyme catalyzed reaction, increase the detectability of the reaction product, and improve the accuracy and stability of detecting the activity of lipoprotein-related phospholipase A2.

Description

technical field [0001] The invention relates to the technical field of clinical medical parameter detection, in particular to a detection reagent, preparation method and application of lipoprotein-related phospholipase A2. Background technique [0002] In recent years, with the improvement of people's living standards, the proportion of nutrients such as sugar and fat that people consume has increased, and the incidence of cardiovascular and cerebrovascular diseases caused by this has increased year by year. Studies have shown that atherosclerosis is the main cause of cardiovascular and cerebrovascular diseases. [0003] The formation of atherosclerosis is due to the promotion of adhesion factors and cytokines in blood vessels. Pro-inflammatory mediators can stimulate the production of adhesion factors and cytokines. The main pro-inflammatory mediators are lysophosphatidylcholine and oxidized free fatty acids. Wait. Medical research results show that lipoprotein-associated...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31G01N31/10
CPCG01N21/31G01N31/10
Inventor 王贤理池万余苗准
Owner 浙江伊利康生物技术有限公司
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