Lipoprotein phosphatidase A2 detection reagent and preparation method and use method thereof

A detection reagent and phospholipase technology, applied in the field of biomedical testing, can solve the problems of difficulty in meeting the needs of industry development, low sensitivity of detection reagents, complicated preparation process, etc., and achieves the advantages of being conducive to widespread clinical use, high cost performance, and simple preparation method. Effect

Inactive Publication Date: 2016-11-09
苏州博源医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection reagents used in these immunological methods have low sensitivity and weak specificity, and the preparation process is complicated. In particular, the detection results are calculated based on the c

Method used

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  • Lipoprotein phosphatidase A2 detection reagent and preparation method and use method thereof
  • Lipoprotein phosphatidase A2 detection reagent and preparation method and use method thereof
  • Lipoprotein phosphatidase A2 detection reagent and preparation method and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 A detection reagent for lipoprotein phospholipase A2

[0043] The detection reagent consists of the following components and their contents, as shown in Table 1:

[0044] Table 1 Components of Lipoprotein Phospholipase A2 Detection Reagent

[0045]

[0046] Wherein, the HEPES buffer (R1) is prepared from the following concentrations of components: 200mmol / L 4-hydroxyethylpiperazineethanesulfonic acid, 10mmol / L 3-[(3-cholesterolaminopropyl)dimethyl Amino]-1-propanesulfonic acid, 10mmol / L sodium n-nonanesulfonate, 5mmol / L ethylenediaminetetraacetic acid, 200mmol / L sodium chloride and the rest of purified water, using 6M HCl to adjust the pH to 7.60± 0.05;

[0047] The citrate buffer (R2a) is prepared from the following components: 20mmol / L citric acid monohydrate, 10mmol / L sodium n-nonanesulfonate and the rest of the purified water, using 6M HCl to adjust the pH to 4.50±0.05;

[0048] The 1-myristoyl-2-(4-p-nitrophenol succinic anhydride) phosphatidylch...

Embodiment 2

[0050] Example 2 A kind of preparation method of lipoprotein phospholipase A2 detection reagent

[0051] Prepare HEPES buffer, citrate buffer, 1-myristoyl-2-(4-p-nitrophenol succinic anhydride) phosphatidylcholine, lipoprotein phospholipase A2 calibrator and lipoprotein phospholipase A2 quality control product;

[0052] The preparation method of described HEPES damping fluid is as follows: take by weighing 4-hydroxyethylpiperazineethanesulfonic acid, 3-[(3-cholesteryl aminopropyl) dimethylamino]-1-propanesulfonic acid, normal Sodium nonanesulfonate, ethylenediaminetetraacetic acid, and sodium chloride were added to purified water, stirred, dissolved, fixed to volume, and adjusted to pH to obtain the product;

[0053] The preparation method of the citrate buffer solution is as follows: Weigh citric acid monohydrate and sodium n-nonanesulfonate in the formula amount, add them into purified water respectively, stir, dissolve, constant volume, and adjust the pH to obtain final ...

Embodiment 3

[0059] Example 3 A method for using a lipoprotein phospholipase A2 detection reagent, comprising the steps of:

[0060] (1) Reagent treatment

[0061] HEPES buffer (R1): stable liquid, after preparation, store at 2-8°C for use;

[0062] Citrate buffer + 1-myristoyl-2-(4-p-nitrophenol succinic anhydride) phosphatidylcholine mixture (R2ab): mix citrate buffer (R2a) with 1-myristoyl- Mix 2-(4-p-nitrophenol succinic anhydride) phosphatidylcholine (R2b) at a volume ratio of 19:1, and the mixture can be stable for two weeks at 2-8°C;

[0063] (2) Prepare samples

[0064] The sample is fresh isolated serum or plasma. Immediately after blood collection, the serum is separated by centrifugation. After centrifugation, the sample can be stored at 2-8°C for 7 days, and stored at -20°C for 2 months;

[0065] (3) Calibration procedure

[0066] Add lipoprotein phospholipase A2 calibrator and HEPES buffer to the automatic biochemical analyzer, mix well, and incubate at 37°C for 4.5 minu...

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Abstract

The invention discloses a lipoprotein phosphatidase A2 detection reagent and a preparation method and a use method thereof. The detection reagent comprises the following components: an 20-200 mmol/L HEPES (hydroxyethyl piperazine ethanesulfonic acid) buffer solution, a 20-100mmol/L citrate buffer solution, a 50 mmol/L 1-myristoyl-2-(4-p-nitrophenol succinic anhydride)phosphatidylcholine solution, a 20-100U/L lipoprotein phospholipase A2 calibration product, and a 20-100U/L lipoprotein phospholipase A2 quality control product. The detection reagent can realize the detection of the lipoprotein phosphatidase A2 by a full automatic biochemical analyzer, can determine the content of the lipoprotein phosphatidase A2 in biological samples in a high-throughput, rapid and accurate mode, and has the advantages of easy operation, high sensitivity, high specificity, accurate results, etc.

Description

technical field [0001] The invention relates to the technical field of biomedical testing, and more specifically relates to a lipoprotein phospholipase A2 detection reagent and a preparation and use method thereof. Background technique [0002] Lipoprotein phospholipase A2 (Lp-PLA2) is an important evaluation index for clinical evaluation and risk prediction of atherosclerosis and coronary heart disease. Lipoprotein phospholipase A2 is a calcium-independent serine lipase. Unlike phospholipases such as cPLA2 and sPLA2, lipoprotein phospholipase A2 is associated with low-density lipoproteins and weakly associated with high-density lipoproteins. Lipoprotein phospholipase A2 is produced by macrophages and other inflammatory cells, and its concentration is higher in the late stage of atherosclerotic lesions than in the early stage. Low-density lipoprotein oxidation is a critical step in the process of atherosclerosis. Lipoprotein phospholipase A2 participates in the degradation...

Claims

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Application Information

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IPC IPC(8): C12Q1/44
CPCC12Q1/44G01N2333/918
Inventor 虞留明张有淘陈薇
Owner 苏州博源医疗科技有限公司
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