Reagent used for measuring pancreatic lipase in serum or plasma, and preparation method and application thereof

A technology of pancreatic lipase and reagents, which is applied in the field of reagents for the determination of pancreatic lipase and its preparation, can solve the problems of false positive detection, increased measured value of pancreatic lipase, and interference with the determination of pancreatic lipase, so as to improve specificity and Accuracy, the effect of reducing the false positive rate

Inactive Publication Date: 2019-04-05
BIOSINO BIO TECH & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Since lipoprotein lipase (LPL), liver lipase (HL) and pancreatic lipase also have the function of lipase, therefore, lipoprotein lipase (LPL) and liver lipase (HL) can also hydrolyze the chromogenic substrate, Increase the measurement value of pancreatic lipase, interfere with the measurement of pancreatic lipase, and lead to false positive detection or clinical misdiagnosis

Method used

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  • Reagent used for measuring pancreatic lipase in serum or plasma, and preparation method and application thereof
  • Reagent used for measuring pancreatic lipase in serum or plasma, and preparation method and application thereof
  • Reagent used for measuring pancreatic lipase in serum or plasma, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The present embodiment provides a reagent for measuring pancreatic lipase, which is composed of reagent 1 and reagent 2, and the specific formula is as follows:

[0053] Reagent 1:

[0054]

[0055] Reagent 2:

[0056]

[0057]

[0058] The present embodiment also provides the preparation method of the assay reagent of above-mentioned pancreatic lipase, specifically as follows:

[0059] Preparation of reagent 1: Add 800mL of deionized water into the container, under magnetic stirring, add 6.057g of Tris, adjust the pH to 8.40 with 6N HCl, then add 0.15g of calcium chloride dihydrate, 2g of sodium taurodeoxycholate, deoxygenated Sodium cholate 2g and co-lipase 1.5mg, until each component is completely dissolved, add deionized water to 1L to obtain reagent 1, its pH is 8.40±0.10.

[0060] Preparation of reagent 2: Add 800 mL of deionized water into the container, and under magnetic stirring, add corresponding amounts of sodium tartrate 1.26 g, tartaric acid 0.5...

Embodiment 2

[0064] The present embodiment provides a reagent for measuring pancreatic lipase, which is composed of reagent 1 and reagent 2, and the specific formula is as follows:

[0065] Reagent 1:

[0066]

[0067] Reagent 2:

[0068]

[0069] The present embodiment also provides the preparation method of the assay reagent of above-mentioned pancreatic lipase, specifically as follows:

[0070] Preparation of reagent 1: Add 800mL of deionized water into the container, add 6.057g of Tris under magnetic stirring, adjust the pH to 8.40 with 6N HCl, then add 0.15g of calcium chloride dihydrate, 1g of sodium lauryl sulfate, and Sodium sulfodeoxycholate 2g, sodium deoxycholate 2g, sphingomyelin 0.1g, protamine sulfate 0.01g and co-lipase 1.5mg, until each component is completely dissolved, add deionized water to 1L, and obtain reagent 1 , and its pH is 8.40±0.10.

[0071] Preparation of reagent 2: Add 800 mL of deionized water into the container, and under magnetic stirring, add corr...

experiment example 1

[0074] Experimental Example 1 Detection Specificity Analysis of Pancrelipase Determination Reagent

[0075] Respectively adopt the pancreatic lipase assay reagent that above-mentioned embodiment 1 and embodiment 2 provide to carry out the mensuration of pancreatic lipase, hepatic lipase, lipoprotein lipase and cholesterol esterase content, use pure water as contrast, analyze pancreatic lipase assay reagent The detection specificity, the specific determination method is as follows:

[0076] 150U / L pancreatic lipase solution, 150U / L liver lipase solution, 150U / L lipoprotein lipase standard solution and 150U / L cholesterol esterase solution were prepared respectively with pure water as solvent. The above solutions were used as samples to be tested.

[0077] Take 4 μL of the above-mentioned sample to be tested and add 250 μL of the reagent 1 of the pancreatic lipase assay reagent of Example 1 and Example 2, and incubate at 37°C for 5 minutes; respectively add 50 μL of the pancreat...

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Abstract

The invention relates to the technical field of biochemical detection, in particular to a reagent used for measuring pancreatic lipase in serum or plasma, and a preparation method and application thereof. The invention provides pancreatic lipase measurement reagent which comprises sodium dodecyl sulfate, protamine sulfate and sphingomyelin. Since the sodium dodecyl sulfate, the protamine sulfate and the sphingomyelin are added into the pancreatic lipase measurement reagent, the interference function of hepatic lipase, lipoprotein lipase and cholesterol esterase in the serum for pancreatic lipase measurement can be obviously lowered, nonspecific lipolysis reaction in the serum or the plasma in a reaction process of the pancreatic lipase measurement can be effectively inhibited, meanwhile, the catalytic activity of the pancreatic lipase can guarantee to be effectively performed, pancreatic lipase measurement specificity and accuracy is improved, and the false positive rate of pancreaticlipase detection is effectively lowered.

Description

technical field [0001] The invention relates to the technical field of biochemical detection, in particular to a reagent for measuring pancreatic lipase in serum or plasma, a preparation method and application thereof. Background technique [0002] Lipase (lipase, Lip), also known as triacylglycerol hydrolase, is a general term for a class of enzymes that can hydrolyze long-chain fatty acid triacylglycerol. According to different synthesis sites, lipase in serum includes: lipoprotein lipase (LPL), liver lipase (HL) and pancreatic lipase (PL). They all belong to lipase family members and have homology. LPL is mainly synthesized in adipocytes, cardiomyocytes, skeletal muscle cells, breast cells and macrophages, and then expressed in the capillary endothelium. LPL mainly acts on TG-rich lipoproteins in plasma, but LPL needs to be combined with apolipoproteins to exert its activity. HL is synthesized by hepatic parenchymal cells and is a key enzyme involved in lipoprotein met...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/77
CPCG01N21/31G01N21/77G01N2021/775
Inventor 张立超
Owner BIOSINO BIO TECH & SCI
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