Preparation method of lipase detection kit

A detection kit, lipase technology, which is used in biological testing, material testing products, color/spectral property measurement, etc. To achieve the effect of increasing the measurable linear range, improving the accuracy and sensitivity, and high accuracy of the results

Inactive Publication Date: 2017-12-08
王贤俊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PNPB method, pH-Stat method, and titration method cannot be used with automatic biochemical analyzers, and are not suitable for clinical diagnosis; most LPS detection kits currently use turbidimetric method, and the defect of this method is that it is highly dependent on the substrate And the accuracy of the reaction is not high, the repeatability is poor, and the linear range is narrow

Method used

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  • Preparation method of lipase detection kit
  • Preparation method of lipase detection kit
  • Preparation method of lipase detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The preparation concentration is the acetic acid-magnesium acetate buffer solution of 80mmol / L, adjusts the pH value to 8.8 as reaction solvent, first adds developer composition, namely 0.1% Tween-80 (Tween-80) and 0.3% fatty alcohol Polyoxyethylene (7) ether, stirring while adding, the rotating speed is controlled at 120r / pm, rotate continuously for 15min, then add the enzyme protection agent composition, that is, the trehalose of 2.5g / L and the sucrose composition of 2.5g / L, After completely dissolving, add the substrate trilaurate 100mmol / L, oxidized coenzyme Ⅰ (NAD + ) 0.2mmol / L, glycerol dehydrogenase (GDH) 0.5KU / L, the enzyme protection agent must be added before adding the enzyme, and finally add the preservative NaN 3 0.6g / L, after being completely mixed, store in a sealed container at 2-8°C.

Embodiment 2

[0028] The preparation concentration is the acetic acid-magnesium acetate buffer solution of 120mmol / L, and the pH value is adjusted to 8.8 as the reaction solvent, and the developer composition is first added, that is, 0.25% Tween-80 (Tween-80) and 0.75% fatty alcohol Polyoxyethylene (7) ether, stirring while adding, the rotating speed is controlled at 120r / pm, rotate continuously for 15min, then add the enzyme protection agent composition, namely the trehalose of 3.5g / L and the sucrose composition of 3.5g / L, After being completely dissolved, 200 mmol / L of glycerol trilaurate, oxidized coenzyme Ⅰ (NAD + ) 0.3mmol / L, glycerol dehydrogenase (GDH) 1.8KU / L, the enzyme protection agent must be added before adding the enzyme, and the preservative NaN should be added at the end 3 0.8g / L, after being completely mixed, store in a sealed container at 2-8°C.

Embodiment 3

[0030] The preparation concentration is the acetic acid-magnesium acetate buffer solution of 160mmol / L, adjusts the pH value to 8.8 as reaction solvent, first adds developer composition, namely 0.4% Tween-80 (Tween-80) and 1.2% fatty alcohol Polyoxyethylene (7) ether, stir while adding, the rotating speed is controlled at 120r / pm, rotate continuously for 15min, then add the enzyme protection agent composition, i.e. the trehalose of 5g / L and the sucrose composition of 5g / L, wait until completely After dissolution, the substrate trilaurate 300mmol / L, oxidized coenzyme Ⅰ (NAD + ) 0.4mmol / L, glycerol dehydrogenase (GDH) 3KU / L, the enzyme protection agent must be added before adding the enzyme, and finally add the preservative NaN 3 1g / L, after being completely mixed, store in a sealed container at 2-8°C.

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Abstract

The invention discloses a preparation method of a lipase detection kit. The preparation method comprises the following steps: preparing a buffer solution with the concentration of 80 to 160mmol/L, adjusting a PH value to be 8.8, and taking the buffer solution as a reaction solvent; adding a developing agent composition, stirring while adding, controlling the rotating speed at 120r/pm and continuously stirring for 15 minutes; then adding a certain amount of enzyme protectant, and respectively adding 100 to 300 mmol/L of laurostearin as a substrate, 0.2 to 0.4 mmol/L of oxidized coenzyme I and 0.5 to 3KU/L of glycerol dehydrogenase (GDH) after the enzyme protectant is completely dissolved, wherein the enzyme protectant is added before the oxidized coenzyme I is added; finally, adding 0.6 to 1g/L of NaN3 as a preservative, and sealing and storing the NaN3 at the temperature of 2 to 8 DEG C after the NaN3 and the solution are uniformly mixed. The method can be used for preparing the lipase detection kit.

Description

technical field [0001] The invention belongs to the field of biological reagents, in particular to a preparation method of a lipase detection kit. Background technique [0002] Lipase (Lipase, glycerol ester hydrolase) belongs to the class of carboxyl ester hydrolase, which can gradually hydrolyze triglycerides into glycerol and fatty acids. It mainly comes from the pancreas, followed by the stomach and small intestine, and can hydrolyze a variety of glycerides containing long-chain (8-18 carbon chain) fatty acids. Lipase is synthesized in the acinar tissue of the pancreas and has high organ specificity. The increase of serum lipase concentration can be detected in the early stage of pancreatitis. For acute abdomen with high fatality rate, timely blood test for serum lipase is of great significance for diagnosis, treatment and prognosis. Early detection of serum lipase can better distinguish acute and non-acute pancreatitis, and its serum expression level has a significant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N33/52
CPCG01N21/31G01N33/52
Inventor 王贤俊张敏
Owner 王贤俊
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