A microtitre plate-based assay for the screening of beta-lactams.
A simple, rapid, sensitive and automatizable method for the detection and quantification of bacterial cell wall inhibitors has been developed. The procedure is characterized by the use of a micro-organism hypersensitive to beta-lactam antibiotics that contains an inducible cytosolic beta-galactosidase; this enzyme is released when the micro-organism cell wall is disrupted by the antibiotic action, and then measured by the use of a chromogenic substrate. The present method allows the detection of beta-lactam traces in other non-beta-lactam antibiotics, and has been successfully applied in the detection of small amounts of beta-lactams in biological fluids such as milk and Actinomycetes fermentation broths. The easy automatization of this method makes it specially suitable for the screening of new antibiotics of natural origin.
Quesada
A R
AR
Departamento de Investigación, Antibióticos S.A., León, Spain.
Cañedo
A
A
Moreno
M A
MA
Fernández-Puentes
J L
JL
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000494:pharmacology; D002417:Cattle; D004353:Drug Evaluation, Preclinical; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology; D005285:Fermentation; D008892:Milk; D010400:Penicillin G / Q000032:analysis; D012680:Sensitivity and Specificity; D013302:Streptomyces / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism; D047090:beta-Lactams / Q000032:analysis* / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D047090:beta-Lactams; D001616:beta-Galactosidase; D010400:Penicillin G