Method and reagent composition for the determination of alanine

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving transferase

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435 4, 435 5, 435 71, 435 25, 435 28, 435810, 436820, C12Q 148, C12Q 126, C12Q 128

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052799441

ABSTRACT:
The enzyme alanine aminotransferase (ALT) is colorimetrically determined as the hydrogen peroxide obtained in the pyruvate hydrolysis reaction catalysed by the enzyme pyruvate oxidase which develops a colour read at 550 nm making use of a reagent composition buffered at pH comprised within 7.0 and 7.5 containing L-alanine, ketoglutaric acid, a source of inorganic phosphorous, the enzyme pyruvate oxidase, a system for revealing the hydrogen peroxide and optionally one or more co-factors which interact in the enzymatic reaction catalysed by the enzyme pyruvate oxidase. It is moreover described a method for the determination of the enzyme alanine aminotransferase making use of said reagent composition and of the surface antigen of the hepatitis B virus (HBsAg), by enzyme-immune assay, in the same serum specimen and in the same well of a microtitration plate. The method is particularly useful for checking the suitability of the blood intended for transfusions since it can be easily carried out with equipments and instruments widely used for the immunoenzymatic screening of infectivity. Furthermore it enables to test in the same analytical assay the two labels of hepatocyte infections necessarily researched in the transfusion centres on biological specimens and to select among the donors the most likely candidates to transmission of the NANB hepatitis virus.

REFERENCES:
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patent: 4707439 (1987-11-01), Seto
patent: 4778757 (1988-10-01), Teshima
patent: 4818688 (1989-04-01), Adamich
patent: 4870026 (1989-09-01), Wands
patent: 5061619 (1991-10-01), Wilson
White, D. O. Medical Virology Academic Press Orlando 1986 pp. 371-373.
IFCC Methods for the Measurement of Catalytic Concentrations of Enzymes Part 3, IFCC Method for Alanine Aminotransferase (L-Alanine: 2-Oxoglutarate Aminotransferase, EC 2.6.1.2), prepared by H. U. Bergmeyer et al., from Clinica Chimica Acta, 105(1980) 147F-154F.

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