Detection of microbial beta-lactamase

Details Detection of microbial beta-lactamase Detection of microbial beta-lactamase

1987-10-29

1990-10-23

Rosen, Sam

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving hydrolase

435 32, 435 29, C12Q 134

Patent

active

049651935

ABSTRACT:
An improved developer for a method for the specific detection of the presence of beta-lactamase from microbial sources is disclosed. The method utilizes a beta-lactam ring-containing substrate whose amide bond is hydrolyzed in the presence of beta-lactamase. A substrate which includes a beta-lactam antibiotic with an acyl side chain containing an alpha-amino group and an alpha-phenyl group or its derivatives is first contacted with an organism thought to produce beta-lactamase or a cell-free beta-lactamase preparation, and, subsequently, it is determined whether the reaction product between the unhydrolyzed substrate, the end products and the organism or the preparation fluoresces. Fluorescence is the indication of .beta.-lactamase activity. The developer comprises the addition of an oxidizing agent (preferably also containing formaldehyde) to the reaction product to enhance the fluorescence. The oxidizing agent is selected from the group consisting of Ag.sup.+, Hg.sup.++, H.sub.2 O.sub.2, I.sub.3.sup.-, IO.sub.4.sup.-, persulfate, Pd.sup.++, p-hydroxymercuribenzoate, and mixtures thereof.

REFERENCES:
patent: 4536334 (1985-08-01), Surayawa et al.
patent: 4668620 (1987-05-01), Armenta et al.
patent: 4740459 (1988-04-01), Chen et al.

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