Compositions for lipase activity determination and method of determining activity

Abstract

To provide lipase activity determination reagents which function by the enzymatic method. The reagents are easy to use in an ordinary clinical examination, have excellent handleability, and are excellent in accuracy and reproducibility. Lipase activity is determined with any of reagents which comprise a low-concentration buffer and a diglyceride dissolved therein. The diglyceride is used as a substrate for lipase, whereby the liquid reagents can have long-term storage stability. One of the reagents converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, pyruvate kinase, lactate dehydrogenase, reduced NAD, ATP, and phosphoenol pyruvate. Another reagent converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, glucose, ADP-dependent hexokinase, glucose-6-phosphate dehydrogenase, oxidized NAD or oxidized NADP, and ATP. A further reagent converts a monoglyceride yielded by a lipase reaction into glycerol with the aid of monoglyceride lipase, and further contains glycerol kinase, glycero-3-phosphate oxidase, peroxidase, and a dye which colors in the presence of hydrogen peroxide.

Description

technical field [0001] The present invention relates to compositions and methods for determining the activity of pancreatic and non-pancreatic lipases, such as hepatic and lipoprotein lipases, in the field of clinical laboratory tests. Background technique [0002] Pancrelipase in blood is an important diagnostic index for pancreatic diseases such as acute pancreatitis, and has been used in routine clinical examinations. [0003] Meanwhile, hepatic lipase or lipoprotein lipase (hereinafter collectively referred to as non-pancreatic lipase) is an important enzyme in the diagnosis of liver diseases or in lipoprotein lipid metabolism. The non-pancreatic lipase plays an important role in the living body as a binder of acidic glycoproteins in the liver or various organs / vascular walls. The non-pancreatic lipase can be made to appear in blood by intravenous injection of heparin or use of heparin in dialysis, etc., but its activity is very low. [0004] Typically, triglycerides a...

AI Technical Summary

Problems solved by technology

[0006] Solutions of diglycerides in nonionic surfactants are known to be unstable and cannot be stored for long periods of time

Therefore, actually, in a reagent or a kit using diglyceride as a substrate, the substrate must be freeze-dried, so a dissolving operation is required before use, making it complicated to use

[0007] Meanwhile, as described above, it is difficult to measure the enzymatic activity of non-pancreatic lipases in blood samples, so in recent years, the determination of the protein levels of the lipases by immunological techniques using monoclonal antibodies has been carried out in the field of clinical laboratory examinations method (non-patent literature 2)

However, the immunological technique used to quantify the protein level of the lipase has the disadvantage of not being able to measure the activity of the enzyme exhibiting the function of the lipase

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