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Testing kit of lipoprotein related phospholipase A2 and its detecting method

A detection method and phospholipase technology, applied in the field of biomedical testing, can solve the problems of affecting the measurement results, limited solubility, etc., and achieve the effects of improving the reaction rate, easy operation, and rapid detection

Inactive Publication Date: 2018-03-13
NINGBO RUI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PAF analogues are lipids with limited solubility in water. In addition, serum enzymes in the sample can also hydrolyze PAF analogues, affecting the measurement results

Method used

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  • Testing kit of lipoprotein related phospholipase A2 and its detecting method
  • Testing kit of lipoprotein related phospholipase A2 and its detecting method
  • Testing kit of lipoprotein related phospholipase A2 and its detecting method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Reagent R1: 50mmol / L carbonic acid buffer, 25mmol / L sodium chloride, 20mmol / L sodium 1-nonanesulfonate, 0.05% sodium azide, pH 7.4.

[0039] Reagent R2A: 50mmol / L carbonic acid buffer, 20mmol / L sodium 1-nonanesulfonate, 20% ethanol, 0.05% sodium azide, pH 4.5.

[0040] Reagent R2B: 50mmol / L phosphate buffer, 10% glycerol, 5mmol / L 1-tetradecanoyl-2-(4-nitrophenylsuccinyl)-3-phosphatidylcholine, 0.05% sodium azide , pH 7.0.

Embodiment 2

[0042] Reagent R1: 20mmol / L phosphate buffer, 15mmol / L disodium edetate, 10mmol / L CHAPS, 0.05% sodium azide, pH 7.0.

[0043] Reagent R2A: 20mmol / L carbonate buffer solution, 10mmol / L sodium 1-nonanesulfonate, 40% glycerol, 0.05% sodium azide, pH 2.7.

[0044] Reagent R2B: 100mmol / L HEPES buffer, 15% ethanol, 10mmol / L 1-tetradecanoyl-2-(4-nitrophenylsuccinyl)-3-phosphatidylcholine, 0.05% sodium azide, pH 7.2.

Embodiment 3

[0046] Reagent R1: 25mmol / L Tris buffer, 10mmol / L potassium chloride, 10mmol / L sodium 1-nonanesulfonate, 0.05% sodium azide, pH 7.5.

[0047] Reagent R2A: 10mmol / L HEPES buffer, 20mmol / L CHAPS, 15% ethanol, 0.05% sodium azide, pH 6.0.

[0048] Reagent R2B: 10mmol / L phosphate buffer, 20% glycerol, 8mmol / L 1-tetradecanoyl-2-(4-nitrophenylsuccinyl)-3-phosphatidylcholine, 0.05% sodium azide , pH 7.0.

[0049] Lp-PLA in the calibrator in embodiment 1-embodiment 3 2 The activity is 336U / L, Lp-PLA in the quality control product 2 Activity is 529U / L, with the solution of 20mmol / L phosphate buffered saline, 150mmol / L sodium chloride, 2% (w / v) BSA, 0.5% (v / v) Tween-20, lipoprotein-associated phospholipase The A2 antigen was prepared into solutions with concentrations of 336U / L and 529U / L, respectively.

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Abstract

The invention belongs to the technical field of biomedicine test, and relates to a testing kit of Lp-PLA2 testing kit and its detecting method. The testing kit comprises reagent R2, reagents R2A and R2B, wherein the reagent R1 is composed of buffer solution, electrolyte, inhibitor, and preservative, and its pH value is 7.0-7.5; the reagent R2A is composed of buffer solution, inhibitor, protectionagent, and preservative, and its pH value is 2.6-6.0; the reagent R2B is composed of buffer solution, Lp-PLA2 substrate, co-solvent, and preservative, and its pH value is 7.0-7.5. the sample and the reagent R1 are evenly mixed and then hatched for 3-5 minutes at 35-40 DEG C; then the reagent R2 prepared by the reagents R2A and R2B is added, and hatched for 60 seconds at 35-40 DEG C; the light absorbency A1 is read; the reagent is hatched for 180 seconds, the light absorbency A2 is read; the differential value of A2 and A2 is calculated; the activity of Lp-PLA2 in the sample is calculated through the calibration curve.

Description

technical field [0001] The invention belongs to the technical field of biomedical testing, and relates to a kit for measuring lipoprotein-related phospholipase A2 and a detection method thereof. Background technique [0002] Lipoprotein-associated phospholipase A2 (Lp-PLA 2 ) is one of the members of the phospholipase A2 superfamily, consisting of 441 amino acids with a relative molecular mass of 45KDa. Because of its ability to degrade platelet-activating factor (PAF) when it was first discovered, it is also called platelet-activating factor acetylhydrolase (platelet-activating factoracetylhydro-lase, PAF-AH). human plasma Lp-PLA 2 It is mainly secreted by macrophages, monocytes, T lymphocytes, and mast cells, and is regulated by various inflammatory factors such as gamma interferon, lipopolysaccharide, and platelet activating factor. In earlier studies, Lp-PLA 2 It can hydrolyze PAF into inactive hemolytic PAF, reduce inflammation and thrombus formation, and it is beli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/92G01N33/53
CPCG01N33/573G01N33/5306G01N33/92G01N2333/916
Inventor 王卫华黄晶晶陈媛张闻周海滨
Owner NINGBO RUI BIO TECH
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