Method of measuring creatine kinase activity

Claim 1

A method for measuring creatine kinase (CK) activity which comprises combining a test sample with the reagents necessary to carry out the following reactions at a pH of about 6 to 7.5: Creatine phosphate+ADP⃡Creatine+ATP (1) ATP+3-Phosphoglycerate⃡ADP+1,3-Diphosphoglycerate (2) 1,3-[Phosphoglycerate]Diphosphoglycerate+NADH⃡Glyceraldehyde-3-phosphate+NAD+Inorganic phosphate (3) wherein one of those reagents is a composition consisting essentially of 0.5 to 50 u/ml of phosphoglycerate kinase (PGK) and 0.5 to 100 u/ml of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the ratio of PGK to GAPDH being about 1:0.2 to 2 and the total amount of PGK and GAPDH in the test being about 0.1 to 30 mg/ml, and as another one of said reagents a buffer solution capable of maintaining said pH of about 6 to 7.5, and optically measuring the rate of reduction of NADH optical absorption.

Claim 2

A method for measuring creatine kinase (CK) activity which comprises combining a test sample with the reagents necessary to carry out the following reactions at a pH of about 6 to 7.5: Creatine phosphate+ADP⃡Creatine+ATP (1) ATP+3-Phosphoglycerate⃡ADP+1,3-Diphosphoglycerate (2) 1,3-[Phosphoglycerate]Diphosphoglycerate+NADH⃡Glyceraldehyde-3-phosphate+NAD+Inorganic phosphate (3) wherein one of those reagents is a composition consisting essentially of 0.5 to 50 u/ml of phosphoglycerate kinase (PGK) and 0.5 to 100 u/ml of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the ratio of PGK to GAPDH being about 1:0.2 to 2 and the total amount of PGK and GAPDH in the test being about 0.1 to 30 mg/ml, and as another one of said reagents a buffer solution capable of maintaining said pH of about 6 to 7.5, and optically measuring the rate of reduction of NADH optical absorption.

Claim 4

A method for measuring creatine kinase (CK) activity which comprises combining a test sample with a reagent composition necessary to carry out the following reactions: Creatine phosphate+ADP⃡Creatine+ATP (1) ATP+3-Phosphoglycerate⃡ADP+1,3-Diphosphoglycerate (2) 1,3-Diphosphoglycerate+NADH⃡Glyceraldehyde-3-phosphate+NAD+Inorganic phosphate (3) wherein the reagent composition for carrying out said reactions consists essentially of: ______________________________________ PGK 0.5-50 μ/ml GAPDH 0.5-100 μ/ml LDH 0-250 μ/ml ADP 0.5-10 mM Creatine phosphate 10-50 mM Magnesium containing salt 5-50 mM 3-Phosphoglycerate 5-50 mM Sulfhydryl compounds 0.1-100 mM Chelating agents 0-10 mM Polyhydroxy compounds 0-0.2 mM AMP 0-25 mM Diadenosine-pentaphosphate 0-100 μM Sterilizers 0-10 mM Buffer (to maintain a pH: 6 to 7.5) 0.05-0.5 M NADH 0.1-0.5 mM Alkaline carbonate or alkaline bicarbonate 0-10 mM Volume fraction of serum in the assay mixture 0.01-0.5, ______________________________________ the ratio of PGK to GAPDH being about 1:0.2 to 2 and the total amount of PGK and GAPDH in the test being about 0.1 to 30 mg/ml, and optically measuring the rate of reduction in NADH optical absorption.

Claim 5

A method for measuring creatine kinase (CK) activity which comprises combining a test sample with a reagent composition necessary to carry out the following reactions: Creatine phosphate+ADP⃡Creatine+ATP (1) ATP+3-Phosphoglycerate⃡ADP+1,3-Diphosphoglycerate (2) 1,3-Diphosphoglycerate+NADH⃡Glyceraldehyde-3-phosphate+NAD+Inorganic phosphate (3) wherein the reagent composition for carrying out said reactions consists essentially of: ______________________________________ PGK 0.5-50 μ/ml GAPDH 0.5-100 μ/ml LDH 0-250 μ/ml ADP 0.5-10 mM Creatine phosphate 10-50 mM Magnesium containing salt 5-50 mM 3-Phosphoglycerate 5-50 mM Sulfhydryl compounds 0.1-100 mM Chelating agents 0-10 mM Polyhydroxy compounds 0-0.2 mM AMP 0-25 mM Diadenosine-pentaphosphate 0-100 μM Sterilizers 0-10 mM Buffer (to maintain a pH: 6 to 7.5) 0.05-0.5 M NADH 0.1-0.5 mM Alkaline carbonate or alkaline bicarbonate 0-10 mM Volume fraction of serum in the assay mixture 0.01-0.5, ______________________________________ the ratio of PGK to GAPDH being about 1:0.2 to 2 and the total amount of PGK and GAPDH in the test being about 0.1 to 30 mg/ml, and optically measuring the rate of reduction in NADH optical absorption.

Claim 6

A method for measuring creatine kinase (CK) activity which comprises combining a test sample with the reagents necessary to carry out the following reactions at a pH of about 6 to 7.5: Creatine phosphate+ADP⃡Creatine+ATP (1) ATP+3-Phosphoglycerate⃡ADP+1,3-Diphosphoglycerate (2) 1,3-Diphosphoglycerate+NADH⃡Glyceraldehyde-3-phosphate+NAD+Inorganic phosphate (3) wherein one of those reagents is a composition consisting essentially of 0.5 to 50 u/ml of phosphoglycerate kinase (PGK), 0.5 to 100 u/ml of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the ratio of PGK to GAPDH being about 1:0.2 to 2 and the total amount of PGK and GAPDH in the test being about 0.1 to 30 mg/ml, and as another one of said reagents a buffer solution capable of maintaining said pH of about 6 to 7.5, and at least one of a stabilizer, a chelating agent, a sterilizer and lactate dehydrogenase (LDH), and optically measuring the rate of reduction of NADH optical absorption.

Claim 7

A method for measuring creatine kinase (CK) activity which comprises combining a test sample with the reagents necessary to carry out the following reactions at a pH of about 6 to 7.5: Creatine phosphate+ADP⃡Creatine+ATP (1) ATP+3-Phosphoglycerate⃡ADP+1,3-Diphosphoglycerate (2) 1,3-Diphosphoglycerate+NADH⃡Glyceraldehyde-3-phosphate+NAD+Inorganic phosphate (3) wherein one of those reagents is a composition consisting essentially of 0.5 to 50 u/ml of phosphoglycerate kinase (PGK), 0.5 to 100 u/ml of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the ratio of PGK to GAPDH being about 1:0.2 to 2 and the total amount of PGK and GAPDH in the test being about 0.1 to 30 mg/ml, and as another one of said reagents a buffer solution capable of maintaining said pH of about 6 to 7.5, and at least one of a stabilizer, a chelating agent, a sterilizer and lactate dehydrogenase (LDH), and optically measuring the rate of reduction of NADH optical absorption.