Highly sensitive enzymatic assay method

Details Highly sensitive enzymatic assay method Highly sensitive enzymatic assay method

1985-07-23

1988-12-13

Warden, Robert J.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving oxidoreductase

435 25, 435 28, 436817, C12Q 126, C12Q 132, C12Q 128

Patent

active

047910577

ABSTRACT:
A highly sensitive quantitative assay method for a component which is 3.beta.-hydroxysteroid or 3-ketosteroid, in a specimen to be assayed, comprising causing this component in the specimen to take part in the cycling reaction ##STR1## and measuring a detectable change in the reaction system. There is thus provided a 3.beta.-hydroxysteroid - 3-ketosteroid cycling reaction using 3.beta.-hydroxysteroid oxidase, which consumes O.sub.2 and generates H.sub.2 O.sub.2 and 3-ketosteroid, with a substrate of 3.beta.-hydroxysteroid, or 3.beta.-hydroxysteroid dehydrogenase, which consumes reduced NAD(P) and generate NAD(P) and 3.beta.-hydroxysteroid, with a substrate of 3-ketosteroid. Example of specimens are specimens which contain 3-hydroxysteroid or 3-ketosteroid, or which liberate or generate such a component. By proceeding at a rate of more than ten cycles per minute and measuring the amount of a detectable change in the reaction, the component in a specimen can easily and sensitively be detected with good accuracy.

REFERENCES:
patent: 4230797 (1980-10-01), Boguslaski et al.
Hurlock et al. (1956) Society of Experimental Biology and Medicine Proceedings, vol. 93, pp. 560-564.
Yamagouchi et al. (1981) Analytical Letters, vol. 14, No. B6, pp. 433-438.
Dolin, 1957 Science, vol. 225, pp. 557-573.
Blaedel et al. (1978), Analytical Chemistry, vol. 50, No. 8, pp. 1026-1032.

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