US 6,686,172 B2Granted

Method of measuring total homocysteine

AssigneeAzwell Inc.

InventorsNaoto Matsuyama, Mina Fukuhara, Masaharu Takayama, Koji Mizuno

Life of this patent

Estimating the term…

Loading patent…

Abstract

The present invention provides a method and a kit for detecting or quantitatively determining homocysteine rapidly and simply and with high sensitivity by oxidizing the residual homocysteine cosubstrate, the produced homocysteine-converting enzyme product or an enzyme reaction product thereof in the presence of an SH reagent to produce hydrogen peroxide and determining the produced hydrogen peroxide by color development using an oxidative color-developing agent. By using the method and kit of the present invention, homocysteine in biological samples, in particular, in body fluids such as blood and urine can be detected and quantitatively determined rapidly and simply and with high sensitivity.

Claims26

Dependent claims

A method for detecting or determining homocysteine in a sample, comprising:

(a) reducing the homocysteine in the sample with a thiol compound,
(b) reacting the reduced homocysteine with a homocysteine-converting enzyme and a homocysteine cosubstrate, thereby producing a homocysteine converting enzyme product, and
(c) detecting or determining the residual homocysteine cosubstrate, the produced homocysteine-converting enzyme product or an enzyme reaction product thereof by (i) oxidizing the residual homocysteine cosubstrate, the produced homocysteine-converting enzyme product or an enzyme reaction product thereof in the presence of an SH reagent to produce hydrogen peroxide and detecting or determining the produced hydrogen peroxide by color development using an oxidative color-developing agent or (ii) reacting the residual homocysteine cosubstrate, the produced homocysteine-converting enzyme product or an enzyme reaction product thereof with a D-amino acid converting enzyme to produce an oxo acid and/or ammonia and detecting or determining the produced oxo acid and/or ammonia.

Dependencies are inferred from the claim text.

Facts

Application: 10/069,847Filed 28 Feb 2002
Priority: JP 2000-19809430 Jun 2000
Published: US 2002/0123088 A15 Sep 2002
Granted: 3 Feb 2004
Term ends: Not estimatedThe source records don’t support an estimate.

Classification8 codes

G01N 33/6815
Assays for specific amino acids: containing sulfur, e.g. cysteine, cystine, methionine, homocysteine
C12Q 1/26
Measuring or testing processes involving enzymes, nucleic acids or microorganisms ; Compositions therefor; Processes of preparing such…
C12Q 1/34
Measuring or testing processes involving enzymes, nucleic acids or microorganisms ; Compositions therefor; Processes of preparing such…
Y10S 435/968
Chemistry: molecular biology and microbiology: high energy substrates, e.g. fluorescent, chemiluminescent, radioactive